Macrocyclic peptides active against the hepatitis c virus

ABSTRACT

Compounds of formula (I): 
     
       
         
         
             
             
         
       
     
     wherein R 1 , R 2 , X, R 3 , D, and the dotted line b are as defined herein; or a pharmaceutically acceptable salt or ester thereof, are useful as inhibitors of the HCV NS3 protease.

This application is a continuation of U.S. application Ser. No.11/039,698, filed Jan. 19, 2005, which claims benefit from U.S.Provisional Application 60/537,863, filed Jan. 21, 2004.

FIELD OF THE INVENTION

The present invention relates to compounds, processes for theirsynthesis, compositions and methods for the treatment of hepatitis Cvirus (HCV) infection. In particular, the present invention providesnovel peptide analogs, pharmaceutical compositions containing suchanalogs and methods for using these analogs in the treatment of HCVinfection.

BACKGROUND OF THE INVENTION

Hepatitis C virus (HCV) is the major etiological agent ofpost-transfusion and community-acquired non-A non-B hepatitis worldwide.It is estimated that over 200 million people worldwide are infected bythe virus. A high percentage of carriers become chronically infected andmany progress to chronic liver disease, so-called chronic hepatitis C.This group is in turn at high risk for serious liver disease such asliver cirrhosis, hepatocellular carcinoma and terminal liver diseaseleading to death.

The mechanism by which HCV establishes viral persistence and causes ahigh rate of chronic liver disease has not been thoroughly elucidated.It is not known how HCV interacts with and evades the host immunesystem. In addition, the roles of cellular and humoral immune responsesin protection against HCV infection and disease have yet to beestablished. Immunoglobulins have been reported for prophylaxis oftransfusion-associated viral hepatitis, however, the Center for DiseaseControl does not presently recommend immunoglobulin treatment for thispurpose. The lack of an effective protective immune response ishampering the development of a vaccine or adequate post-exposureprophylaxis measures, so in the near-term, hopes are firmly pinned onantiviral interventions.

Various clinical studies have been conducted with the goal ofidentifying pharmaceutical agents capable of effectively treating HCVinfection in patients afflicted with chronic hepatitis C. These studieshave involved the use of interferon-alpha, alone and in combination withother antiviral agents. Such studies have shown that a substantialnumber of the participants do not respond to these therapies, and ofthose that do respond favorably, a large proportion were found torelapse after termination of treatment.

Until recently, interferon (IFN) was the only available therapy ofproven benefit approved in the clinic for patients with chronichepatitis C. However the sustained response rate is low, and interferontreatment also induces severe side-effects (i.e. retinopathy,thyroiditis, acute pancreatitis, depression) that diminish the qualityof life of treated patients. Recently, interferon in combination withribavirin has been approved for patients non-responsive to IFN alone.However, the side effects caused by IFN are not alleviated with thiscombination therapy. Pegylated forms of interferons such as PEG-Intron®and Pegasys® can apparently partially address these deleteriousside-effects but antiviral drugs still remain the avenue of choice fororal treatment of HCV.

Therefore, a need exists for the development of effective antiviralagents for treatment of HCV infection that overcome the limitations ofexisting pharmaceutical therapies.

HCV is an enveloped positive strand RNA virus in the Flaviviridaefamily. The single strand HCV RNA genome is approximately 9500nucleotides in length and has a single open reading frame (ORF) encodinga single large polyprotein of about 3000 amino acids. In infected cells,this polyprotein is cleaved at multiple sites by cellular and viralproteases to produce the structural and non-structural (NS) proteins. Inthe case of HCV, the generation of mature nonstructural proteins (NS2,NS3, NS4A, NS4B, NS5A, and NS5B) is effected by two viral proteases. Thefirst one, as yet poorly characterized, cleaves at the NS2—NS3 junction(henceforth referred to as NS2/3 protease); the second one is a serineprotease contained within the N-terminal region of NS3 (NS3 protease)and mediates all the subsequent cleavages downstream of NS3, both incis, at the NS3—NS4A cleavage site, and in trans, for the remainingNS4A-NS4B, NS4B-NS5A, NS5A-NS5B sites. The NS4A protein appears to servemultiple functions, acting as a cofactor for the NS3 protease andpossibly assisting in the membrane localization of NS3 and other viralreplicase components. The complex formation of the NS3 protease withNS4A seems necessary to the processing events, enhancing the proteolyticefficiency at all of the sites. The NS3 protein also exhibits nucleosidetriphosphatase and RNA helicase activities. NS5B is a RNA-dependent RNApolymerase that is involved in the replication of HCV.

A general strategy for the development of antiviral agents is toinactivate virally encoded enzymes that are essential for thereplication of the virus. In a two day clinical trial, it has been shownthat the HCV NS3 protease inhibitor BILN 2061 is effective in rapidlyreducing viral loads in patients infected with the hepatitis C virus(Nature (2003) 426, p. 186-189), thus providing proof of principle ofthe clinical antiviral activity of HCV NS3 protease inhibitors.

The NS3 protease has been found to potentially have an additional impactby blocking the IFN-mediated cellular antiviral activity in the infectedcell (Foy et al., Science, 17 Apr. 2003). This lends credence to ahypothesis that the NS3/NS4A protease may represent a dual therapeutictarget, the inhibition of which may both block viral replication andrestore Interferon response of HCV infected cells.

Macrocyclic compounds which inhibit the HCV NS3 protease have beendescribed in WO 00/59929 (U.S. Pat. No. 6,608,027), WO 03/053349, WO03/064455 and WO 2004/037855.

The present invention now provides novel compounds that are inhibitoryto the NS3 protease. Furthermore, compounds being active in cell cultureare provided.

An advantage of one aspect of the present invention resides in the factthat compounds according to this invention specifically inhibit the NS3protease and do not show significant inhibitory activity against otherserine proteases such as human leukocyte Elastase (HLE), porcinepancreatic Elastase (PPE), or bovine pancreatic chymotrypsin, orcysteine proteases such as human liver Cathepsin B (Cat B).

SUMMARY OF THE INVENTION

Included in the scope of the invention are compounds of formula (I):

wherein R¹ is hydroxy or NHSO₂R¹¹ wherein R¹¹ is (C₁₋₆)alkyl,(C₂₋₆)alkenyl, (C₃₋₇)cycloalkyl, (C₃₋₇)cycloalkyl-(C₁₋₆)alkyl-, aryl,Het, aryl-(C₁₋₄)alkyl-, or Het-(C₁₋₄)alkyl-;

-   -   a) said (C₁₋₆)alkyl, (C₂₋₆)alkenyl, aryl, Het,        (C₃₋₇)cycloalkyl-(C₁₋₆)alkyl-, aryl-(C₁₋₄)alkyl-, and        Het-(C₁₋₄)alkyl- optionally being substituted with one, two or        three substituents each independently selected from halogen,        hydroxy, cyano, nitro, (C₁₋₆)alkyl, (C₁₋₆)haloalkyl,        —O—(C₁₋₆)alkyl, —O—(C₁₋₆)haloalkyl, —O-aryl, —C(═O)—(C₁₋₆)alkyl,        —C(═O)—NH₂, —C(═O)—NH(C₁₋₄)alkyl, —C(═O)—N((C₁₋₄)alkyl)₂, —NH₂,        —NH(C₁₋₄)alkyl and —N((C₁₋₄)alkyl)₂; and    -   b) said (C₃₋₇)cycloalkyl being optionally substituted with one        or more substituents each independently selected from nitro,        halogen, hydroxy, cyano, —O—(C₁₋₆)alkyl, (C₂₋₄)alkenyl,        —O—(C₁₋₆)haloalkyl, —NH₂, —NH(C₁₋₄)alkyl, —N((C₁₋₄)alkyl)₂,        tri(C₁₋₆)alkylsilyl, R⁴¹, —C(═O)—R⁴¹—C(═O)OR⁴¹, —C(═O)N(R⁴²)R⁴¹,        —SO₂R⁴¹, and —OC(═O)—R⁴¹;        -   wherein R⁴¹ in each case is independently selected from:        -   i) H, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, Het, or            aryl-(C₁₋₄)alkyl-O—;        -   ii) aryl or aryloxy, each of which being optionally            substituted with (C₁₋₆)alkyl; and        -   iii) (C₁₋₈)alkyl optionally substituted with one or more            substituents each independently selected from            —O—(C₁₋₆)alkyl, hydroxy, halogen, (C₂₋₁₀)alkenyl,            (C₂₋₁₀)alkynyl, (C₃₋₇)cycloalkyl, (C₄₋₇)cycloalkenyl, aryl,            Het, aryloxy, and aryl-(C₁₋₄)alkyl-O—, wherein each of said            aryl and aryloxy is optionally substituted with (C₁₋₆)alkyl;            and        -   R⁴² is selected from H and (C₁₋₆)alkyl; or

-   R¹¹ is —N(R^(11a))(R^(11b)), wherein R^(11a) and R^(11b) are each    independently selected from H, (C₁₋₆)alkyl, (C₃₋₇)cycloalkyl,    (C₃₋₇)cycloalkyl-(C₁₋₆)alkyl-, aryl, aryl-(C₁₋₆)alkyl-, Het and    Het-(C₁₋₄)alkyl-; wherein said (C₁₋₆)alkyl, (C₃₋₇)cycloalkyl,    (C₃₋₇)cycloalkyl-(C₁₋₆)alkyl-, aryl, aryl-(C₁₋₆)alkyl-, Het and    Het-(C₁₋₄)alkyl- are each optionally substituted with one or more    substituents each independently selected from halogen, (C₁₋₆)alkyl,    hydroxy, cyano, nitro, (C₁₋₆)haloalkyl, —O—(C₁₋₆)alkyl,    —O—(C₁₋₆)haloalkyl, —NH₂, —NH(C₁₋₄)alkyl, —N((C₁₋₄)alkyl)₂,    —C(═O)—NH₂, —C(═O)—NH(C₁₋₄)alkyl, —C(═O)—N((C₁₋₄)alkyl)₂,    —C(═O)—(C₁₋₆)alkyl, —COOH, and —COO(C₁₋₆)alkyl; or    -   R^(11a) and R^(11b) are linked, together with the nitrogen to        which they are bonded, to form a 3- to 7-membered monocyclic        saturated or unsaturated heterocycle optionally fused to at        least one other cycle to form a heteropolycycle, said        heterocycle and heteropolycycle optionally containing from one        to three further heteroatoms each independently selected from N,        S and O, and being optionally substituted with one or more        substituents each independently selected from halogen,        (C₁₋₆)alkyl, hydroxy, cyano, nitro, (C₁₋₆)haloalkyl,        —O—(C₁₋₆)alkyl, —O—(C₁₋₆)haloalkyl, —NH₂, —NH(C₁₋₄)alkyl,        —N((C₁₋₄)alkyl)₂, —C(═O)—NH₂, —C(═O)—NH(C₁₋₄)alkyl,        —C(═O)—N((C₁₋₄)alkyl)₂, —C(═O)—(C₁₋₆)alkyl, —COOH, and        —COO(C₁₋₆)alkyl;

-   R² is a group of formula:

wherein

-   -   R²⁰ is H, OH, halogen, or Y¹—R^(20a) wherein Y¹ is a bond, O, S,        or NR^(20b) and    -   wherein:        -   R^(20a) is selected from the group consisting of:            (C₁₋₈)alkyl, (C₁₋₆)alkyl-C≡N, (C₂₋₈)alkenyl, (C₂₋₈)alkynyl            and (C₃₋₇)cycloalkyl, each of said alkyl, alkenyl, alkynyl            and cycloalkyl being optionally substituted with one, two or            three substituents, each independently selected from:            -   halogen, (C₁₋₆)alkyl optionally substituted with                —O—(C₁₋₆)alkyl or —O—(C₃₋₆)cycloalkyl, (C₃₋₇)cycloalkyl,                —O—(C₁₋₆)alkyl, Het, —O—(C₃₋₆)cycloalkyl, —NH₂,                —NH(C₁₋₄)alkyl and —N((C₁₋₄)alkyl)₂; and        -   R^(20b) is H, (C₁₋₆)alkyl or (C₃₋₆)cycloalkyl;        -   and W is N; and the dotted line “a” is a double bond; or    -   R²⁰ is oxo, and W is NR²³ wherein R²³ is H, (C₁₋₆)alkyl,        (C₂₋₆)alkenyl or (C₂₋₆)alkynyl; and the dotted line “a” is a        single bond;    -   R²¹ is halogen or Y²—R^(21a), wherein Y² is a bond, O, S, SO or        SO₂, and R^(21a) is (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynyl,        (C₃₋₇)cycloalkyl or (C₃₋₇)cycloalkyl-(C₁₋₆)alkyl-;    -   R²² is H, —OH, —O—(C₁₋₄)alkyl, —NH₂, —NH(C₁₋₄)alkyl or        —N((C₁₋₄)alkyl)₂;        or R² is a group of formula:

wherein

-   -   R²⁰, W and the dotted line “a” are as defined above;    -   R²⁴ is H or R²¹ as defined above; and    -   R²⁵ is H or (C₁₋₆)alkyl;

-   X is O or NH;

-   R³ is (C₁₋₁₀)alkyl, (C₃₋₇)cycloalkyl or    (C₃₋₇)cycloalkyl-(C₁₋₄)alkyl-,    -   a) wherein the cycloalkyl and cycloalkyl-alkyl- may be mono-,        di- or tri-substituted with (C₁₋₃)alkyl;    -   b) wherein the alkyl, cycloalkyl and cycloalkyl-alkyl- may be        mono- or di-substituted with substituents each independently        selected from hydroxy and O—(C₁₋₆)alkyl;    -   c) wherein each alkyl group may be mono-, di- or tri-substituted        with halogen; and    -   d) wherein in each cycloalkyl group being 5-, 6- or 7-membered,        one or two —CH₂-groups not being directly linked to each other        may be replaced by —O— such that the O-atom is linked to the        group X via at least two C-atoms;

-   D is a 3 to 8 atom saturated or unsaturated alkylene chain; and    the dotted line “b” is a single bond or a double bond;    wherein Het as used herein is defined as a 3- to 7-membered    heterocycle having 1 to 4 heteroatoms each independently selected    from O, N and S, which may be saturated, unsaturated or aromatic,    and which is optionally fused to at least one other cycle to form a    4- to 14-membered heteropolycycle having wherever possible 1 to 5    heteroatoms, each independently selected from O, N and S, said    heteropolycycle being saturated, unsaturated or aromatic;    or a pharmaceutically acceptable salt or ester thereof;    with the proviso that    when R² is a group of formula

and

-   W is N; and the dotted line “a” is a double bond; and-   R²⁰ is H, halogen, or Y¹—R^(20a), wherein Y¹ is O and R^(20a) is    (C₁₋₆)alkyl or (C₃₋₆)cycloalkyl; or Y¹ is a bond and R^(20a) is    (C₁₋₆)alkyl; and-   R²¹ is halogen or Y² R^(21a), wherein Y² is O and R^(21a) is    (C₁₋₆)alkyl or (C₃₋₆)cycloalkyl; and-   R²² is H; and-   R³ is (C₁₋₆)alkyl optionally substituted with halo, or R³ is    —(CH₂)_(p)—(C₃₋₇)cycloalkyl wherein p is 0-4, or R³ is a    tetrahydrofuran ring linked through the C3 or C4 position of the    ring;-   then R¹ is not NHSO₂R¹¹ wherein R¹¹ is (C₁₋₆)alkyl or unsubstituted    (C₃₋₇)cycloalkyl.

Included within the scope of this invention is a pharmaceuticalcomposition comprising an anti-hepatitis C virally effective amount of acompound of formula (I), or a pharmaceutically acceptable salt or esterthereof, in admixture with at least one pharmaceutically acceptablecarrier medium or auxiliary agent.

According to a further aspect of this embodiment the pharmaceuticalcomposition as defined above further comprises a therapeuticallyeffective amount of at least one other antiviral agent.

Another important aspect of the invention involves a method of treatingor preventing a hepatitis C viral infection in a mammal by administeringto the mammal an anti-hepatitis C virally effective amount of a compoundof formula (I), a pharmaceutically acceptable salt or ester thereof, ora composition as described above, alone or in combination with at leastone other antiviral agent, administered together or separately.

Also within the scope of this invention is the use of a compound offormula (I), or a pharmaceutically acceptable salt or ester thereof, asdescribed herein, for the manufacture of a medicament for the treatmentor prevention of hepatitis C viral infection in a mammal.

A further aspect of the invention provides the use of a compound offormula (I), or a pharmaceutically acceptable salt or ester thereof, asdescribed herein, in combination with at least one other antiviralagent, for the manufacture of a medicament for the treatment orprevention of hepatitis C viral infection in a mammal.

Still another aspect of this invention relates to a method of inhibitingthe replication of hepatitis C virus by exposing the virus to ahepatitis C viral NS3 protease inhibiting amount of the compound offormula (I) according to this invention, or a pharmaceuticallyacceptable salt thereof.

Yet a further aspect of this invention provides the use of a compound offormula (I), or a pharmaceutically acceptable salt or ester thereof, asdescribed herein, to inhibit the replication of hepatitis C virus.

An additional aspect of this invention refers to an article ofmanufacture comprising a composition effective to treat an HCV infectionor to inhibit the NS3 protease of HCV and packaging material comprisinga label which indicates that the composition can be used to treatinfection by the hepatitis C virus, wherein said composition comprises acompound of formula (I) according to this invention or apharmaceutically acceptable salt thereof.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS Definitions

As used herein, the following definitions apply unless otherwise noted:

With reference to the instances where (R) or (S) is used to designatethe absolute configuration of a substituent or asymmetric center of acompound of formula (I), the designation is done in the context of thewhole compound and not in the context of the substituent or asymmetriccenter alone.

The designation “P1, P2, and P3” as used herein refer to the position ofthe amino acid residues starting from the C-terminus end of the peptideanalogs and extending towards the N-terminus (i.e. P1 refers to position1 from the C-terminus, P2: second position from the C-terminus, etc.)(see Berger A. & Schechter I., Transactions of the Royal Society Londonseries B257, 249-264 (1970)).

As used herein the term “(1R,2S)-vinyl-ACCA” refers to a compound offormula:

namely, (1R,2S) 1-amino-2-ethenylcyclopropylcarboxylic acid.

The term “(C_(x-n))alkyl” as used herein, either alone or in combinationwith another substituent, means acyclic, straight or branched chainalkyl substituents containing from x to n carbon atoms, wherein x is 0(i.e. the alkyl group is absent) or an integer and n is an integer.“(C₁₋₆)alkyl” (or “lower alkyl”) includes, for example, methyl, ethyl,n-propyl, n-butyl, 1-methylethyl (iso-propyl), 1-methylpropyl,2-methylpropyl, 1,1-dimethylethyl (tert-butyl), pentyl and hexyl. Theabbreviation Me denotes a methyl group and, Et denotes ethyl.

The term “(C_(2-n))alkenyl” as used herein, either alone or incombination with another substituent, means acyclic, straight orbranched chain alkyl substituents containing from 2 to n carbon atoms,wherein n is an integer, at least two of which are linked by a doublebond (within the carbon chain or terminal). Examples of (C_(2-n))alkenylinclude, but are not limited to, ethenyl (vinyl), 1-propenyl,2-propenyl, and 1-butenyl. The cis and trans isomers, and mixturesthereof, of the (C_(2-n))alkenyl radical can be encompassed by the term.A (C_(2-n))alkenyl radical may be substituted on any of the carbon atomsthereof which would otherwise bear a hydrogen atom.

The term “(C_(2-n))alkynyl” as used herein, either alone or incombination with another substituent, means acyclic, straight orbranched chain alkyl substituents containing from 2 to n carbon atoms,wherein n is an integer, at least two of which are linked by a triplebond (within the carbon chain or terminal). Examples of such radicalsinclude, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, and1-butynyl. A (C_(2-n))alkynyl radical may be substituted on any of thecarbon atoms thereof which would otherwise bear a hydrogen atom.

The term “(C_(3-m))cycloalkyl” as used herein, either alone or incombination with another substituent, means a cycloalkyl substituentcontaining from 3 to m carbon atoms, wherein m is an integer andincludes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl and cycloheptyl.

The terms “(C_(x-n))alkyl-(C_(3-m))cycloalkyl” or“(C_(3-m))cycloalkyl-(C_(x-n))alkyl-” as used herein interchangeablymean an alkylene radical containing from x to n carbon atoms to which acycloalkyl radical containing from 3 to m carbon atoms, wherein x is 0(i.e. the alkylene radical is absent) or an integer and n and m are eachindependently an integer, is directly linked; for example,cyclopropylmethyl, cyclopentylethyl, cyclohexylmethyl,1-cyclohexylethyl, 2-cyclohexylethyl and cycloheptylpropyl.

The term “aryl” or “(C_(6 or 10))aryl” as used herein interchangeably,either alone or in combination with another radical, means either anaromatic monocyclic group containing 6 carbon atoms or an aromaticbicyclic group containing 10 carbon atoms. For example, aryl includesphenyl, 1-naphthyl and 2-naphthyl.

As used herein interchangeably, the terms“(C_(x-n))alkyl-(C_(6 or 10))aryl” or“(C_(6 or 10))aryl-(C_(x-n))alkyl-” means an alkyl radical as definedabove to which an aryl substituent as defined above is bonded. Examplesof (C_(6 or 10))aryl-(C₁₋₃)alkyl- are benzyl (phenylmethyl),1-phenylethyl, 2-phenylethyl and phenylpropyl.

The term “O—(C_(1-n))alkyl” or “(C_(1-n))alkoxy” as used hereininterchangeably, either alone or in combination with another radical,means the radical —O—(C_(1-n))alkyl wherein alkyl is as defined abovecontaining from 1 to n carbon atoms, and includes methoxy, ethoxy,propoxy, 1-methylethoxy, butoxy and 1,1-dimethylethoxy. The latterradical is known commonly as tert-butoxy. When an —O—(C_(1-n))alkylgroup is substituted, it is understood to be substituted on the(C_(1-n))alkyl portion thereof.

The term “S—(C_(1-n))alkyl” or “(C_(1-n))alkylthio” as used hereininterchangeably, either alone or in combination with another radical,means the radical —S—(C_(1-n))alkyl wherein alkyl is as defined abovecontaining from 1 to n carbon atoms, and includes methylthio, ethylthio,propylthio, 1-methylethylthio, butylthio and 1,1-dimethylethylthio. Whenan —S—(C_(1-n))alkyl group is substituted, it is understood to besubstituted on the (C_(1-n))alkyl portion thereof.

The term “halo” or “halogen” as used herein interchangeably, means ahalogen substituent selected from fluoro, chloro, bromo or iodo.

The terms “(C₁₋₆)haloalkyl” or “lower haloalkyl”, as used hereininterchangeably, mean an alkyl radical containing one to six carbonatoms wherein one or more hydrogen atoms are replaced by a halogen atom(including but not limited to trifluoromethyl).

The term “oxo” as used herein means an oxygen atom attached as asubstituent by a double bond (═O).

As used herein, the term “Het” defines a 3- to 7-membered heterocyclehaving 1 to 4 heteroatoms each independently selected from O, N and S,which may be saturated, unsaturated or aromatic, and which is optionallyfused to at least one other cycle to form a 4- to 14-memberedheteropolycycle having wherever possible 1 to 5 heteroatoms, eachindependently selected from O, N and S, said heteropolycycle beingsaturated, unsaturated or aromatic, unless specified otherwise.

As used herein the term “heteroatom” means O, S or N.

As used herein, the term “heterocycle”, either alone or in combinationwith another radical, means a monovalent radical derived by removal of ahydrogen from a three- to seven-membered saturated or unsaturated(including aromatic) heterocycle containing from one to four heteroatomseach independently selected from nitrogen, oxygen and sulfur. Examplesof such heterocycles include, but are not limited to, azetidine,pyrrolidine, tetrahydrofuran, thiazolidine, pyrrole, thiophene,hydantoin, diazepine, 1H-imidazole, isoxazole, thiazole, tetrazole,piperidine, piperazine, homopiperidine, homopiperazine, 1,4-dioxane,4-morpholine, 4-thiomorpholine, pyridine, pyridine-N-oxide orpyrimidine, or the following heterocycles:

As used herein, the term “heteropolycycle” either alone or incombination with another radical, means a heterocycle as defined abovefused to one or more other cycle, be it a heterocycle or any othercycle. Examples of such heteropolycycles include, but are not limitedto, indole, benzimidazole, thiazolo[4,5-b]-pyridine, quinoline,isoquinoline, or coumarin, or the following:

Although generally covered under the term “Het”, the term “heteroaryl”as used herein precisely defines an unsaturated heterocycle for whichthe double bonds form an aromatic system. Suitable examples of“heteroaryl” include but are not limited to: quinoline, indole,pyridine,

The term “pharmaceutically acceptable ester” as used herein, eitheralone or in combination with another substituent, means esters of thecompound of formula (I) in which any of the carboxyl functions of themolecule, but preferably the carboxy terminus, is replaced by analkoxycarbonyl function:

in which the R moiety of the ester is selected from alkyl (e.g. methyl,ethyl, n-propyl, tert-butyl, n-butyl); alkoxyalkyl (e.g. methoxymethyl);alkoxyacyl (e.g. acetoxymethyl); aralkyl (e.g. benzyl); aryloxyalkyl(e.g. phenoxymethyl); aryl (e.g. phenyl), optionally substituted withhalogen, (C₁₋₄)alkyl or (C₁₋₄)alkoxy. Other suitable prodrug esters canbe found in Design of prodrugs, Bundgaard, H. Ed. Elsevier (1985). Suchpharmaceutically acceptable esters are usually hydrolyzed in vivo wheninjected in a mammal and transformed into the acid form of the compoundof formula (I). With regard to the esters described above, unlessotherwise specified, any alkyl moiety present advantageously contains 1to 16 carbon atoms, particularly 1 to 6 carbon atoms. Any aryl moietypresent in such esters advantageously comprises a phenyl group. Inparticular the esters may be a (C₁₋₁₆)alkyl ester, an unsubstitutedbenzyl ester or a benzyl ester substituted with at least one halogen,(C₁₋₆)alkyl, (C₁₋₆)alkoxy, nitro or trifluoromethyl.

The term “pharmaceutically acceptable salt” means a salt of a compoundof formula (I) which is, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response, and the like,commensurate with a reasonable benefit/risk ratio, generally water oroil-soluble or dispersible, and effective for their intended use. Theterm includes pharmaceutically-acceptable acid addition salts andpharmaceutically-acceptable base addition salts. Lists of suitable saltsare found in, e.g., S. M. Birge et al., J. Pharm. Sci., 1977, 66, pp.1-19.

The term “pharmaceutically-acceptable acid addition salt” means thosesalts which retain the biological effectiveness and properties of thefree bases and which are not biologically or otherwise undesirable,formed with inorganic acids such as hydrochloric acid, hydrobromic acid,sulfuric acid, sulfamic acid, nitric acid, phosphoric acid, and thelike, and organic acids such as acetic acid, trifluoroacetic acid,adipic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoicacid, butyric acid, camphoric acid, camphorsulfonic acid, cinnamic acid,citric acid, digluconic acid, ethanesulfonic acid, glutamic acid,glycolic acid, glycerophosphoric acid, hemisulfic acid, hexanoic acid,formic acid, fumaric acid, 2-hydroxyethanesulfonic acid (isethionicacid), lactic acid, hydroxymaleic acid, malic acid, malonic acid,mandelic acid, mesitylenesulfonic acid, methanesulfonic acid,naphthalenesulfonic acid, nicotinic acid, 2-naphthalenesulfonic acid,oxalic acid, pamoic acid, pectinic acid, phenylacetic acid,3-phenylpropionic acid, pivalic acid, propionic acid, pyruvic acid,salicylic acid, stearic acid, succinic acid, sulfanilic acid, tartaricacid, p-toluenesulfonic acid, undecanoic acid, and the like.

The term “pharmaceutically-acceptable base addition salt” means thosesalts which retain the biological effectiveness and properties of thefree acids and which are not biologically or otherwise undesirable,formed with inorganic bases such as ammonia or hydroxide, carbonate, orbicarbonate of ammonium or a metal cation such as sodium, potassium,lithium, calcium, magnesium, iron, zinc, copper, manganese, aluminum,and the like. Particularly preferred are the ammonium, potassium,sodium, calcium, and magnesium salts. Salts derived frompharmaceutically-acceptable organic nontoxic bases include salts ofprimary, secondary, and tertiary amines, quaternary amine compounds,substituted amines including naturally occurring substituted amines,cyclic amines and basic ion-exchange resins, such as methylamine,dimethylamine, trimethylamine, ethylamine, diethylamine, triethylamine,isopropylamine, tripropylamine, tributylamine, ethanolamine,diethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol,dicyclohexylamine, lysine, arginine, histidine, caffeine, hydrabamine,choline, betaine, ethylenediamine, glucosamine, methylglucamine,theobromine, purines, piperazine, piperidine, N-ethylpiperidine,tetramethylammonium compounds, tetraethylammonium compounds, pyridine,N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine,dicyclohexylamine, dibenzylamine, N,N-dibenzylphenethylamine,1-ephenamine, N,N′-dibenzylethylenediamine, polyamine resins, and thelike. Particularly preferred organic nontoxic bases are isopropylamine,diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline,and caffeine.

The term “mammal” as it is used herein is meant to encompass humans, aswell as non-human mammals which are susceptible to infection byhepatitis C virus including domestic animals, such as cows, pigs,horses, dogs and cats, and non-domestic animals.

The term “antiviral agent” as used herein means an agent (compound orbiological) that is effective to inhibit the formation and/orreplication of a virus in a mammal. This includes agents that interferewith either host or viral mechanisms necessary for the formation and/orreplication of a virus in a mammal. Such agents can be selected from:another anti-HCV agent, HIV inhibitor, HAV inhibitor and HBV inhibitor.Antiviral agents include, for example, ribavirin, amantadine, VX-497(merimepodib, Vertex Pharmaceuticals), VX-498 (Vertex Pharmaceuticals),Levovirin, Viramidine, Ceplene (maxamine), XTL-001 and XTL-002 (XTLBiopharmaceuticals).

The term “other anti-HCV agent” as used herein means those agents thatare effective for diminishing or preventing the progression of hepatitisC related symptoms of disease. Such agents can be selected from:immunomodulatory agents, inhibitors of HCV NS3 protease, inhibitors ofHCV polymerase or inhibitors of another target in the HCV life cycle.

The term “immunomodulatory agent” as used herein means those agents(compounds or biologicals) that are effective to enhance or potentiatethe immune system response in a mammal. Immunomodulatory agents include,for example, class I interferons (such as α-, β-, δ-, ω- andτ-interferons, consensus interferons and asialo-interferons), class IIinterferons (such as γ-interferons) and pegylated interferons.

The term “inhibitor of HCV NS3 protease” as used herein means an agent(compound or biological) that is effective to inhibit the function ofHCV NS3 protease in a mammal. Inhibitors of HCV NS3 protease include,for example, those compounds described in WO 99/07733, WO 99/07734, WO00/09543, WO 00/09558, WO 00/59929, WO 03/064416, WO 03/064455, WO03/064456, WO 2004/037855 and co-pending patent application Ser. Nos.10/850,101 and 60/504,839, herein incorporated by reference in theirentirety (all by Boehringer Ingelheim), WO 02/060926, WO 03/053349, WO03/099316, WO 03/099274, WO 2004/032827 and US 2004/0077551 (all byBMS), WO 2004/072243 (Enanta) and the Vertex clinical candidateidentified as VX-950.

The term “inhibitor of HCV polymerase” as used herein means an agent(compound or biological) that is effective to inhibit the function of anHCV polymerase in a mammal. This includes, but is not limited to,non-nucleoside and nucleoside inhibitors of HCV NS5B polymerase.

Examples of inhibitors of HCV polymerase include but are not limited tothose compounds described in: WO 02/04425 (Boehringer Ingelheim) WO03/007945 (Boehringer Ingelheim), WO 03/010140 (Boehringer Ingelheim),WO 03/010141 (Boehringer Ingelheim), WO 2004/064925 (BoehringerIngelheim), WO 2004/065367 (Boehringer Ingelheim), WO 2004/087714(IRBM), WO 03/101993 (Neogenesis), WO 03/026587 (BMS), WO 03/000254(Japan Tobacco), and WO 01/47883 (Japan Tobacco), and the clinicalcandidates JTK-003 (Japan Tobacco), HCV 086 (ViroPharma/Wyeth), R-803(Rigel) and NM 283 (Idenix/Novartis).

The term “inhibitor of another target in the HCV life cycle” as usedherein means an agent (compound or biological) that is effective toinhibit the formation and/or replication of HCV in a mammal other thanby inhibiting the function of the HCV NS3 protease. This includes agentsthat interfere with either host or HCV viral mechanisms necessary forthe formation and/or replication of HCV in a mammal. Inhibitors ofanother target in the HCV life cycle include, for example, agents thatinhibit a target selected from a helicase, a NS2/3 protease and aninternal ribosome entry site (IRES). Specific examples of inhibitors ofanother target in the HCV life cycle include ISIS-14803 (ISISPharmaceuticals).

The term “HIV inhibitor” as used herein means an agent (compound orbiological) that is effective to inhibit the formation and/orreplication of HIV in a mammal. This includes agents that interfere witheither host or viral mechanisms necessary for the formation and/orreplication of HIV in a mammal. HIV inhibitors include, for example,nucleoside inhibitors, non-nucleoside inhibitors, protease inhibitors,fusion inhibitors and integrase inhibitors.

The term “HAV inhibitor” as used herein means an agent (compound orbiological) that is effective to inhibit the formation and/orreplication of HAV in a mammal. This includes agents that interfere witheither host or viral mechanisms necessary for the formation and/orreplication of HAV in a mammal. HAV inhibitors include Hepatitis Avaccines, for example, Havrix® (GlaxoSmithKline), VAQTA® (Merck) andAvaxim® (Aventis Pasteur).

The term “HBV inhibitor” as used herein means an agent (compound orbiological) that is effective to inhibit the formation and/orreplication of HBV in a mammal. This includes agents that interfere witheither host or viral mechanisms necessary for the formation and/orreplication of HBV in a mammal. HBV inhibitors include, for example,agents that inhibit HBV viral DNA polymerase or HBV vaccines. Specificexamples of HBV inhibitors include Lamivudine (Epivir-HBV®), AdefovirDipivoxil, Entecavir, FTC (Coviracil®), DAPD (DXG), L-FMAU (Clevudine®),AM365 (Amrad), Ldt (Telbivudine), monoval-LdC (Valtorcitabine),ACH-126,443 (L-Fd4C) (Achillion), MCC478 (Eli Lilly), Racivir (RCV),Fluoro-L and D nucleosides, Robustaflavone, ICN 2001-3 (ICN), Bam 205(Novelos), XTL-001 (XTL), Imino-Sugars (Nonyl-DNJ) (Synergy), HepBzyme;and immunomodulator products such as: interferon alpha 2b, HE2000(Hollis-Eden), Theradigm (Epimmune), EHT899 (Enzo Biochem), Thymosinalpha-1 (Zadaxin®), HBV DNA vaccine (PowderJect), HBV DNA vaccine(Jefferon Center), HBV antigen (OraGen), BayHep B® (Bayer), Nabi-HB®(Nabi) and Anti-hepatitis B (Cangene); and HBV vaccine products such asthe following: Engerix B, Recombivax HB, GenHevac B, Hepacare, Bio-HepB, TwinRix, Comvax, Hexavac.

The term “class I interferon” as used herein means an interferonselected from a group of interferons that all bind to receptor type I.This includes both naturally and synthetically produced class Iinterferons. Examples of class I interferons include α-, β-, δ-, ω- andτ-interferons, consensus interferons, asialo-interferons and pegylatedforms thereof.

The term “class II interferon” as used herein means an interferonselected from a group of interferons that all bind to receptor type II.Examples of class II interferons include γ-interferons.

Specific preferred examples of some of these agents are listed below:

-   -   antiviral agents: ribavirin and amantadine;    -   immunomodulatory agents: class I interferons, class II        interferons and pegylated forms thereof;    -   HCV polymerase inhibitors: nucleoside analogs and        non-nucleosides;    -   inhibitor of another target in the HCV life cycle that inhibits        a target selected from: NS3 helicase, NS2/3 protease or internal        ribosome entry site (IRES);    -   HIV inhibitors: nucleoside inhibitors, non-nucleoside        inhibitors, protease inhibitors, fusion inhibitors and integrase        inhibitors; or    -   HBV inhibitors: agents that inhibit viral DNA polymerase or is        an HBV vaccine.

As discussed above, combination therapy is contemplated wherein acompound of formula (I), or a pharmaceutically acceptable salt thereof,is co-administered with at least one additional agent selected from: anantiviral agent, an immunomodulatory agent, an inhibitor of HCVpolymerase, another inhibitor of HCV NS3 protease, an inhibitor ofanother target in the HCV life cycle, an HIV inhibitor, an HAV inhibitorand an HBV inhibitor. Examples of such agents are provided in theDefinitions section above. These additional agents may be combined withthe compounds of this invention to create a single pharmaceutical dosageform. Alternatively these additional agents may be separatelyadministered to the patient as part of a multiple dosage form, forexample, using a kit. Such additional agents may be administered to thepatient prior to, concurrently with, or following the administration ofa compound of formula (I), or a pharmaceutically acceptable saltthereof.

As used herein, the term “treatment” means the administration of acompound or composition according to the present invention to alleviateor eliminate symptoms of the hepatitis C disease and/or to reduce viralload in a patient.

As used herein, the term “prevention” means the administration of acompound or composition according to the present invention post-exposureof the individual to the virus but before the appearance of symptoms ofthe disease, and/or prior to the detection of the virus in the blood toprevent the appearance of symptoms of the disease and/or to prevent thevirus from reaching detectible levels in the blood.

The following sign - - - or → are used interchangeably in sub-formulasto indicate the bond which is connected to the rest of the molecule asdefined.

Preferred Embodiments

In the following preferred embodiments, groups and substituents of thecompounds according to this invention are described in detail.

Included in the preferred embodiments of the invention are compounds offormula (I) wherein:

R¹:

According to one preferred embodiment of the present invention, R¹ ishydroxy.

According to an alternative preferred embodiment of the presentinvention, R¹ is NHSO₂R¹¹; wherein

-   -   R¹¹ is selected from methyl, ethyl, propyl, 1-methylethyl,        butyl, 1-methylpropyl, 2-methylpropyl, 1,1-dimethylethyl,        ethenyl, 1-propenyl, 2-propenyl, cyclopropylmethyl,        cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, phenyl,        naphthyl, Het, phenylmethyl, naphthylmethyl and Het-methyl;    -   a) each of which optionally being mono-, di- or tri-substituted        with substituents each independently selected from fluorine and        methyl; and    -   b) each of which optionally being mono- or disubstituted with        substituents each independently selected from hydroxy,        trifluoromethyl, methoxy, phenoxy and trifluoromethoxy; and    -   c) each of which optionally being monosubstituted with a        substituent selected from chlorine, bromine, cyano, nitro,        —CO—NH₂, —CO—NHCH₃, —CO—N(CH₃)₂, —NH₂, —NH(CH₃) and —N(CH₃)₂;    -   wherein Het is selected from thienyl, furyl, thiazolyl,        benzothiazolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridinyl,        pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl,        3H-indolyl, indolyl, quinolinyl, isoquinolinyl,        tetrahydrothienyl, tetrahydrofuryl, thiadiazolyl, isoxazolyl,        benzothienyl, piperidinyl, piperazinyl, morpholinyl, triazolyl,        tetrazolyl, and

According to another alternative preferred embodiment, R¹ is NHSO₂R¹¹;wherein

-   -   R¹¹ is selected from cyclopropyl, cyclobutyl, cyclopentyl and        cyclohexyl;    -   a) each of which optionally being mono-, di- or tri-substituted        with fluorine; and    -   b) each of which optionally being mono- or disubstituted with        substituents selected from hydroxy, methoxy and        trifluoromethoxy; and    -   c) each of which optionally being monosubstituted with a        substituent selected from chlorine, bromine, cyano, nitro,        —CO—NH₂, —CO—NHCH₃, —CO—N(CH₃)₂, —NH₂, —NH(CH₃) and —N(CH₃)₂;        and    -   d) each of which being optionally substituted with one or more        (C₁₋₈)alkyl, wherein each (C₁₋₈)alkyl is independently        optionally substituted with one or more substituents each        independently selected from —O—(C₁₋₆)alkyl, hydroxy, halogen,        (C₂₋₁₀)alkenyl, (C₂₋₁₀)alkynyl, (C₃₋₇)cycloalkyl,        (C₄₋₇)cycloalkenyl, aryl, aryloxy, and aryl-(C₁₋₄)alkyl-O—,        wherein each of said aryl and aryloxy is optionally substituted        with (C₁₋₆)alkyl.

According to yet another alternative preferred embodiment, R¹ isNHSO₂R¹¹ wherein R¹¹ is —N(R^(11a))(R^(11b)),

-   -   wherein R^(11a) and R^(11b) are each independently selected from        H, methyl, ethyl, propyl, 1-methylethyl, cyclopropyl,        cyclobutyl, cyclopentyl, cyclohexyl, phenyl and phenylmethyl;        wherein said methyl, ethyl, propyl, 1-methylethyl, cyclopropyl,        cyclobutyl, cyclopentyl, cyclohexyl, phenyl and phenylmethyl are        optionally substituted with one or more substituents each        independently selected from halogen, (C₁₋₄)alkyl, hydroxy,        cyano, O—(C₁₋₄)alkyl, —NH₂, —NH(C₁₋₄)alkyl, —N((C₁₋₄)alkyl)₂,        —CO—NH₂, —CO—NH(C₁₋₄)alkyl, —CO—N((C₁₋₄)alkyl)₂, —COOH, and        —COO(C₁₋₄)alkyl; or    -   R^(11a) and R^(11b) are linked, together with the nitrogen to        which they are bonded, to form a 3- 4-, 5- or 6-membered        monocyclic saturated or unsaturated heterocycle, optionally        containing from one to three further heteroatoms each        independently selected from N, S and O, and optionally        substituted with one, two or three substituents each        independently selected from halogen, (C₁₋₄)alkyl, hydroxy,        cyano, O—(C₁₋₄)alkyl, —NH₂, —NH(C₁₋₄)alkyl, —N((C₁₋₄)alkyl)₂,        —CO—NH₂, —CO—NH(C₁₋₄)alkyl, —CO—N((C₁₋₄)alkyl)₂, —COOH, and        —COO(C₁₋₄)alkyl.

According to this alternative preferred embodiment, R^(11a) and R^(11b)are more preferably each independently selected from methyl, ethyl,propyl, 1-methylethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,phenyl and phenylmethyl; wherein said methyl, ethyl, propyl,1-methylethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyland phenylmethyl are optionally substituted with one or moresubstituents each independently selected from halogen, (C₁₋₄)alkyl,hydroxy, cyano, O—(C₁₋₄)alkyl, —NH₂, —NH(C₁₋₄)alkyl, —N((C₁₋₄)alkyl)₂,—CO—NH₂, —CO—NH(C₁₋₄)alkyl, —CO—N((C₁₋₄)alkyl)₂, —COOH, and—COO(C₁₋₄)alkyl; or

R^(11a) and R^(11b) are linked, together with the nitrogen to which theyare bonded, to form a 3- 4-, 5- or 6-membered monocyclic saturated orunsaturated heterocycle, optionally containing from one to three furtherheteroatoms each independently selected from N, S and O, and optionallysubstituted with one, two or three substituents each independentlyselected from halogen, (C₁₋₄)alkyl, hydroxy, cyano, O—(C₁₋₄)alkyl, —NH₂,—NH(C₁₋₄)alkyl, —N((C₁₋₄)alkyl)₂, —CO—NH₂, —CO—NH(C₁₋₄)alkyl,—CO—N((C₁₋₄)alkyl)₂, —COOH, and —COO(C₁₋₄)alkyl.

Therefore, more preferably, when R¹ is NHSO₂R¹¹, the group R¹¹ isselected from methyl, ethyl, 1-methylethyl, propyl, ethenyl,cyclopropyl, cyclobutyl, cyclopentyl, phenyl,

and —N(CH₃)₂; wherein said phenyl is optionally monosubstituted withhalogen and wherein said cyclopropyl is optionally substituted at the1-position with methyl, ethyl, propyl or butyl, each of said methyl,ethyl, propyl and butyl being optionally further substituted withphenyl, (C₃₋₆)cycloalkyl, (C₂₋₆)alkenyl or (C₁₋₄)alkoxy.

Most preferably, R¹¹ is methyl, cyclopropyl, phenyl,

or —N(CH₃)₂. R²

According to a preferred embodiment, R² is a group of formula:

whereinW, R²⁰, R²¹, R²² and the dotted line “a” are as defined herein.

According to an alternative preferred embodiment, R² is a group offormula:

whereinW, R²⁰, R²⁴, R²⁵ and the dotted line “a” are as defined herein.

W and R²⁰:

Preferably, when R² is a group of formula

R²⁰ is oxo; W is NR²³ wherein R²³ is preferably Me, Et, —CH₂CH═CH₂ or H;and the dotted line “a” is a single bond.

Alternatively preferably, when R² is a group of formula

W is N, the dotted line “a” is a double bond; and R²⁰ is H, OH, halogen,or Y¹—R^(20a) wherein

-   -   Y¹ is a bond, O, S, or NR^(20b);    -   R^(20a) is selected from the group consisting of: (C₁₋₈)alkyl,        (C₂₋₈)alkenyl, (C₂₋₈)alkynyl and (C₃₋₇)cycloalkyl, each of said        alkyl, alkenyl, alkynyl and cycloalkyl being optionally        substituted with one, two or three substituents, each        independently selected from:        -   halogen, (C₁₋₆)alkyl, (C₃₋₇)cycloalkyl, Het, —O—(C₁₋₆)alkyl,            —O—(C₃₋₆)cycloalkyl, —NH₂, —NH(C₁₋₄)alkyl and            —N((C₁₋₄)alkyl)₂; and    -   R^(20b) is H, (C₁₋₆)alkyl or (C₃₋₆)cycloalkyl.

More preferably, when W is N and the dotted line “a” is a double bond,R²⁰ is H, (C₁₋₆)alkyl, OH, —O—(C₁₋₆)alkyl, —S—(C₁₋₆)alkyl,—(CH₂)₀₋₄—CH═CH₂, —(CH₂)₀₋₄—C≡CH, —O—(CH₂)₀₋₄—CH═CH₂, —O—(CH₂)₀₋₄—C═CH,—O—(CH₂)₁₋₄—OMe; —O—(CH₂)₁₋₄—N(Me)₂; —O—(CH₂)₁₋₄-Het;—S—(CH₂)₀₋₄—CH═CH₂, —S—(CH₂)₀₋₄—C≡CH, —S—(CH₂)₁₋₄—OMe;—S—(CH₂)₁₋₄—N(Me)₂, —S—(CH₂)₁₋₄-Het; (C₃₋₆)cycloalkyl,—O—(C₃₋₆)cycloalkyl, —O—(C₁₋₆)alkyl-(C₃₋₆)cycloalkyl,—S—(C₃₋₆)cycloalkyl, or —S—(C₁₋₆)alkyl-(C₃₋₆)cycloalkyl; wherein Het is5- or 6-membered monocyclic heteroaryl containing from one to threeheteroatoms each independently selected from N, O and S;

-   -   each of said (C₁₋₆)alkyl, —O—(C₁₋₆)alkyl, —S—(C₁₋₆)alkyl,        —(CH₂)₀₋₄—CH═CH₂, —(CH₂)₀₋₄—C≡CH, —O—(CH₂)₀₋₄—CH═CH₂,        —O—(CH₂)₀₋₄—C≡CH, —S—(CH₂)₀₋₄—CH═CH₂, —S—(CH₂)₀₋₄—C≡CH,        (C₃₋₆)cycloalkyl, —O—(C₃₋₆)cycloalkyl, and —S—(C₃₋₆)cycloalkyl        being optionally substituted with one, two or three        substituents, each independently selected from (C₁₋₄)alkyl,        —O—(C₁₋₄)alkyl, and halo;        or R²⁰ is NR^(20a)R^(20b) wherein R^(20a) is (C₁₋₄)alkyl, and        R^(20b) is H, (C₁₋₄)alkyl or (C₃₋₅)cycloalkyl.

Even more preferably R²⁰ is H, methyl, ethyl, propyl, 1-methylethyl,butyl, 2-methylpropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,—CH═CH₂, —C≡CH, O-methyl, O-ethyl, O-propyl, O—CH(CH₃)₂, O-cyclopropyl,O-cyclobutyl, O-cyclopentyl, O-cyclohexyl, O—CH₂CH₂CF₃, —O—CH═CH₂,—O—CH₂—CH═CH₂, O—C≡CH, —O—CH₂—C═CH, —O—CH₂—C≡CCH₃, —O—CH₂—CH₂—OMe,—O—CH₂—CH₂—N(Me)₂, S-methyl, S-ethyl, S-propyl, S—CH(CH₃)₂,S-cyclopropyl, S-cyclobutyl, S-cyclopentyl, S-cyclohexyl, —S—CH═CH₂,—S—CH₂—CH═CH₂, S—C≡CH, —S—CH₂—C≡CH, —S—CH₂—CH₂-OMe, —S—CH₂—CH₂—N(Me)₂,

Most preferably R²⁰ is H, methyl, ethyl, 1-methylethyl, —C≡CH, O-methyl,O-ethyl, O-propyl, O—CH(CH₃)₂, O-cyclopentyl, O—CH₂CH₂CF₃, —O—CH═CH₂,—O—CH₂—CH═CH₂, —O—CH₂—C≡CH, —O—CH₂—C≡CCH₃, —O—CH₂CH₂OMe,—O—CH₂CH₂N(Me)₂, S-methyl, S-ethyl, S-propyl, S—CH(CH₃)₂,

Preferably, when R² is a group of formula

W is N; the dotted line “a” is a double bond; and R²⁰ is H, OH, halogen,or Y¹—R^(20a) wherein

-   -   Y¹ is a bond, O, S, or NR^(20b);    -   R^(20a) is selected from the group consisting of: (C₁₋₈)alkyl,        (C₂₋₈)alkenyl, (C₂₋₈)alkynyl and (C₃₋₇)cycloalkyl, each of said        alkyl, alkenyl, alkynyl and cycloalkyl being optionally        substituted with one, two or three substituents, each        independently selected from:        -   halogen, (C₁₋₆)alkyl, (C₃₋₇)cycloalkyl, Het, —O—(C₁₋₆)alkyl,            —O—(C₃₋₆)cycloalkyl, —NH₂, —NH(C₁₋₄)alkyl and            —N((C₁₋₄)alkyl)₂; and    -   R^(20b) is H, (C₁₋₆)alkyl or (C₃₋₆)cycloalkyl.

More preferably, R²⁰ is Y¹—R^(20a), wherein Y¹ is O and R^(20a) is(C₁₋₈)alkyl. Most preferably, R²⁰ is —O—CH₂CH₃.

R²¹:

R²¹ is preferably selected from: fluorine, chlorine, bromine, —CH₃,—CH₂CH₃, —CH₂CH₂CH₃, —OCH₃, —OCH₂CH₃, —OCH₂CH₂CH₃, —SCH₃, —SCH₂CH₃,—SCH₂CH₂CH₃, —(SO)CH₃, —(SO)CH₂CH₃, —(SO)CH₂CH₂CH₃, —(SO₂)CH₃,—(SO₂)CH₂CH₃, —(SO₂)CH₂CH₂CH₃,

and —C≡CH.

More preferably, R²¹ is selected from: fluorine, chlorine, bromine,—CH₃, —OCH₃, —SCH₃, —SCH₂CH₃, —(SO)CH₃, —(SO₂)CH₃, —(SO₂)CH₂CH₃,

and —C≡CH. R²².

R²² is preferably selected from: H, —OH, —OCH₃, —OCH₂CH₃, —OCH₂CH₂CH₃,—NHCH₃, —N(CH₃)₂, —N(CH₃)CH₂CH₃ and —N(CH₃)CH₂CH₂CH₃.

More preferably R²² is selected from H, —OCH₃ and —N(CH₃)₂.

Most preferably, R²² is H or —OCH₃.

R²⁴.

Preferably, R²⁴ is H or (C₁₋₆)alkyl. Most preferably, R²⁴ is H or CH₃.

R²⁵:

Most preferably, R²⁵ is H.

X:

According to one embodiment of this invention X is O.

According to another embodiment of this invention X is NH.

Preferably, X is O.

R³.

With respect to compounds of formula (I) as defined above, R³ ispreferably selected from (C₂₋₈)alkyl, (C₃₋₇)cycloalkyl and(C₃₋₇)cycloalkyl-(C₁₋₃)alkyl-,

-   a) wherein said cycloalkyl and cycloalkyl-alkyl- may be mono-, di-    or tri-substituted with (C₁₋₃)alkyl; and-   b) wherein said alkyl, cycloalkyl and cycloalkyl-alkyl- may be mono-    or di-substituted with substituents each independently selected from    hydroxy and O—(C₁₋₄)alkyl; and-   c) wherein each of said alkyl groups may be mono-, di- or    tri-substituted with fluorine or mono-substituted with chlorine or    bromine; and-   d) wherein in each of said cycloalkyl groups being 5-, 6- or    7-membered, one or two —CH₂-groups not being directly linked to each    other may be replaced by —O— such that the O-atom is linked to the    group X via at least two C-atoms.    R³ is more preferably selected from ethyl, 1-methylethyl,    1,1-dimethylethyl, propyl, 1-methylpropyl, 2-methylpropyl,    1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl,    1,1,2-trimethylpropyl, 1,2,2-trimethylpropyl, 1-ethylpropyl,    1-ethyl-2-methylpropyl, 1-(1-methylethyl)-2-methylpropyl,    1-ethyl-2,2-dimethylpropyl, butyl, 1-methylbutyl, 2-methylbutyl,    3-methylbutyl, 1,2-dimethylbutyl, 1,1-dimethylbutyl,    1,3-dimethylbutyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl,    3,3-dimethylbutyl, 1,2,2-trimethylbutyl, 1,2,3-trimethylbutyl,    2,3,3-trimethylbutyl and 2,2,3-trimethylbutyl, whereby these alkyl    groups may be substituted with chlorine or bromine, or with 1, 2 or    3 fluorine substituents. Examples of preferred fluorinated alkyl    groups include, but are not limited to, 2-fluoroethyl,    3-fluoropropyl and 3,3,3-trifluoropropyl.

Alternatively more preferably, R³ is cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl or is selected from the following formulas,wherein one or two CH₂-groups of a cycloalkyl group is replaced byoxygen:

From the above list, cycloalkyl and cycloalkyl-alkyl- groups optionallycomprising 1 or 2 O-atoms are optionally substituted with 1, 2 or 3methyl groups. Especially those cycloalkyl groups, optionally comprising1 or 2 O-atoms, are preferred, wherein the α-C-atom is substituted withmethyl.

Further examples of preferred substituted cyclic groups are

Even more preferred meanings of R³ are cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, and a group selected from:

Most preferably R³ is selected from cyclopentyl,

D:

Preferred embodiments of the present invention include compounds offormula (I), wherein linker D is a 3 to 8 atom saturated or unsaturatedalkylene chain. More preferably, linker D is a 5 carbon atom chain.

Dotted Line “b”:

Preferably the dotted line “b” is a single bond or a double bond. Morepreferably, the dotted line “b” is a single bond or a double bond in theZ (cis) configuration.

Therefore, according to one embodiment of this invention, compounds areprovided of formula (Ia):

wherein R¹ is hydroxy or NHSO₂R¹¹ wherein R¹¹ is (C₁₋₆)alkyl,(C₃₋₇)cycloalkyl, (C_(6 or 10))aryl, Het, (C₃₋₇)cycloalkyl-(C₁₋₆)alkyl-,(C_(6 or 10))aryl-(C₁₋₄)alkyl- or Het-(C₁₋₄)alkyl-, all of which beingoptionally mono-, di- or tri-substituted with substituents selectedfrom:

-   -   halogen, hydroxy, cyano, nitro, (C₁₋₆)alkyl, (C₁₋₆)haloalkyl,        O—(C₁₋₆)alkyl, —O—(C₁₋₄)haloalkyl, —C(O)—(C₁₋₆)alkyl, —C(O)—NH₂,        —C(O)—NH(C₁₋₄)alkyl, —C(O)—N((C₁₋₄)alkyl)₂, —NH₂, —NH(C₁₋₄)alkyl        and —N((C₁₋₄)alkyl)₂;        or R¹¹ is —NR^(11a)R^(11b) wherein R^(11a) is H or (C₁₋₆)alkyl,        and R^(11b) is H, (C₁₋₆)alkyl, (C₃₋₆)cycloalkyl,        (C_(6 or 10))aryl, Het, (C₃₋₆)cycloalkyl-(C₁₋₄)alkyl-,        (C_(6 or 10))aryl-(C₁₋₄)alkyl- or Het-(C₁₋₄)alkyl-, or R^(11a)        and R^(11b) are linked to each other to form a 3 to 7-membered        nitrogen-containing ring optionally containing one or two        further heteroatoms selected from: O, S or N, all of said        R^(11a) and R^(11b) being optionally substituted with:    -   halogen, hydroxy, cyano, nitro, (C₁₋₆)alkyl, (C₁₋₆)haloalkyl,        O—(C₁₋₆)alkyl, —O—(C₁₋₄)haloalkyl, —C(O)—(C₁₋₆)alkyl, —C(O)—NH₂,        —C(O)—NH(C₁₋₄)alkyl, —C(O)—N((C₁₋₄)alkyl)₂, —NH₂, —NH(C₁₋₄)alkyl        and —N((C₁₋₄)alkyl)₂;

-   R²⁰ is H, OH, halogen, or Y¹—R^(20a) wherein Y¹ is a bond, O, S, or    NR^(20b) wherein:    -   R^(20a) is selected from the group consisting of: (C₁₋₈)alkyl,        (C₁₋₆)alkyl-C≡N, (C₂₋₈)alkenyl, (C₂₋₈)alkynyl, all of said        alkyl, alkenyl and alkynyl being optionally mono- or        di-substituted with:        -   halogen, (C₁₋₆)alkyl, —O—(C₁₋₆)alkyl,            (C₁₋₄)alkyl-O—(C₁₋₆)alkyl, —O—(C₃₋₆)cycloalkyl,            (C₁₋₄)alkyl-O—(C₃₋₆)cycloalkyl, amino, (C₁₋₆)alkylamino, or            di((C₁₋₆)alkyl)amino; and    -   R^(20b) is H, (C₁₋₆)alkyl or (C₃₋₆)cycloalkyl;

-   and W is N; and the dotted line “a” is a double bond; or

-   R²⁰ is oxo, and W is NR²³ wherein R²³ is H, (C₁₋₆)alkyl,    (C₂₋₆)alkenyl, (C₂₋₆)alkynyl; and the dotted line “a” is a single    bond;

-   R²¹ is halogen, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynyl,    —O—(C₁₋₆)alkyl, —O—(C₂₋₆)alkenyl, —O—(C₂₋₆)alkynyl, —S—(C₁₋₆)alkyl,    —S—(C₂₋₆)alkenyl, and —S—(C₂₋₆)alkynyl, wherein the sulfur is in any    oxidized state;

-   R²² is H, —OH, —O—(C₁₋₄)alkyl, —NH₂, —NH(C₁₋₄)alkyl or    —N((C₁₋₄)alkyl)₂;

-   X is O or NH;

-   R³ is (C₁₋₁₀)alkyl, (C₃₋₇)cycloalkyl, (C₃₋₇)cycloalkyl-(C₁₋₄)alkyl-,    -   a) wherein the cycloalkyl and cycloalkyl-alkyl- may be mono-,        di- or tri-substituted with (C₁₋₃)alkyl;    -   b) wherein the alkyl, cycloalkyl and cycloalkyl-alkyl- may be        mono- or di-substituted with substituents selected from hydroxy        and O—(C₁₋₆)alkyl;    -   c) wherein all the alkyl groups may be mono-, di- or        tri-substituted with halogen; and    -   d) wherein in the cycloalkyl groups, being 5-, 6- or 7-membered,        one or two —CH₂-groups not being directly linked to each other        may be replaced by —O—;

-   D is a 3 to 8 atom saturated or unsaturated alkylene chain; and    the dotted line “b” is a single bond or a double bond;    or a pharmaceutically acceptable salt or ester thereof.

In an alternative preferred embodiment are compounds of formula (I)wherein

-   R¹ is hydroxy or NHSO₂R¹¹; wherein-   R¹¹ is selected from methyl, ethyl, propyl, 1-methylethyl, butyl,    1-methylpropyl, 2-methylpropyl, 1,1-dimethylethyl, ethenyl,    1-propenyl, 2-propenyl, cyclopropylmethyl, cyclobutylmethyl,    cyclopentylmethyl, cyclohexylmethyl, phenyl, naphthyl, Het,    phenylmethyl, naphthylmethyl and Het-methyl;    -   a) each of which optionally being mono-, di- or tri-substituted        with substituents each independently selected from fluorine and        methyl; and    -   b) each of which optionally being mono- or disubstituted with        substituents each independently selected from hydroxy,        trifluoromethyl, methoxy, phenoxy and trifluoromethoxy; and    -   c) each of which optionally being monosubstituted with a        substituent selected from chlorine, bromine, cyano, nitro,        —CO—NH₂, —CO—NHCH₃, —CO—N(CH₃)₂, —NH₂, —NH(CH₃) and —N(CH₃)₂;    -   wherein Het is selected from thienyl, furyl, thiazolyl,        benzothiazolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridinyl,        pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl,        3H-indolyl, indolyl, quinolinyl, isoquinolinyl,        tetrahydrothienyl, tetrahydrofuryl, thiadiazolyl, isoxazolyl,        benzothienyl, piperidinyl, piperazinyl, morpholinyl, triazolyl,        tetrazolyl, and

or

-   R¹¹ is selected from cyclopropyl, cyclobutyl, cyclopentyl and    cyclohexyl;    -   a) each of which optionally being mono-, di- or tri-substituted        with fluorine; and    -   b) each of which optionally being mono- or disubstituted with        substituents selected from hydroxy, methoxy and        trifluoromethoxy; and    -   c) each of which optionally being monosubstituted with a        substituent selected from chlorine, bromine, cyano, nitro,        —CO—NH₂, —CO—NHCH₃, —CO—N(CH₃)₂, —NH₂, —NH(CH₃) and —N(CH₃)₂;        and    -   d) each of which being optionally substituted with one or more        (C₁₋₈)alkyl, wherein each (C₁₋₈)alkyl is independently        optionally substituted with one or more substituents each        independently selected from —O—(C₁₋₆)alkyl, hydroxy, halogen,        (C₂₋₁₀)alkenyl, (C₂₋₁₀)alkynyl, (C₃₋₇)cycloalkyl,        (C₄₋₇)cycloalkenyl, aryl, aryloxy, and aryl-(C₁₋₄)alkyl-O—,        wherein each of said aryl and aryloxy is optionally substituted        with (C₁₋₆)alkyl; or-   R¹¹ is N(R^(11a))(R^(11b)), wherein    -   R^(11a) and R^(11b) are each independently selected from H,        methyl, ethyl, propyl, 1-methylethyl, cyclopropyl, cyclobutyl,        cyclopentyl, cyclohexyl, phenyl and phenylmethyl; wherein said        methyl, ethyl, propyl, 1-methylethyl, cyclopropyl, cyclobutyl,        cyclopentyl, cyclohexyl, phenyl and phenylmethyl are optionally        substituted with one or more substituents each independently        selected from halogen, (C₁₋₄)alkyl, hydroxy, cyano,        O—(C₁₋₄)alkyl, —NH₂, —NH(C₁₋₄)alkyl, —N((C₁₋₄)alkyl)₂, —CO—NH₂,        —CO—NH(C₁₋₄)alkyl, —CO—N((C₁₋₄)alkyl)₂, —COOH, and        —COO(C₁₋₄)alkyl; or    -   R^(11a) and R^(11b) are linked, together with the nitrogen to        which they are bonded, to form a 3- 4-, 5- or 6-membered        monocyclic saturated or unsaturated heterocycle, optionally        containing from one to three further heteroatoms each        independently selected from N, S and O, and optionally        substituted with one, two or three substituents each        independently selected from halogen, (C₁₋₄)alkyl, hydroxy,        cyano, O—(C₁₋₄)alkyl, —NH₂, —NH(C₁₋₄)alkyl, —N((C₁₋₄)alkyl)₂,        —CO—NH₂, —CO—NH(C₁₋₄)alkyl, —CO—N((C₁₋₄)alkyl)₂, —COOH, and        —COO(C₁₋₄)alkyl; and-   R² is a group of formula

wherein

-   -   R²⁰ is oxo, W is NR²³ wherein R²³ is Me, Et, —CH₂CH═CH₂ or H and        the dotted line “a” is a single bond; or    -   W is N, the dotted line “a” is a double bond; and R²⁰ is H, OH,        halogen, or Y¹—R^(20a)        -   wherein        -   Y¹ is a bond, O, S, or NR^(20b);        -   R^(20a) is selected from the group consisting of:            (C₁₋₈)alkyl, (C₂₋₈)alkenyl, (C₂₋₈)alkynyl and            (C₃₋₇)cycloalkyl, each of said alkyl, alkenyl, alkynyl and            cycloalkyl being optionally substituted with one, two or            three substituents, each independently selected from:            -   halogen, (C₁₋₆)alkyl, (C₃₋₇)cycloalkyl, Het,                —O—(C₁₋₆)alkyl, —O—(C₃₋₆)cycloalkyl, —NH₂,                —NH(C₁₋₄)alkyl and —N((C₁₋₄)alkyl)₂; and        -   R^(20b) is H, (C₁₋₆)alkyl or (C₃₋₆)cycloalkyl; and    -   R²¹ is selected from: fluorine, chlorine, bromine, —CH₃,        —CH₂CH₃, —CH₂CH₂CH₃, —OCH₃, —OCH₂CH₃, —OCH₂CH₂CH₃, —SCH₃,        —SCH₂CH₃, —SCH₂CH₂CH₃, —(SO)CH₃, —(SO)CH₂CH₃, —(SO)CH₂CH₂CH₃,        —(SO₂)CH₃, —(SO₂)CH₂CH₃, —(SO₂)CH₂CH₂CH₃,

and —C≡CH; and

-   -   R²² is selected from: H, —OH, —OCH₃, —OCH₂CH₃, —OCH₂CH₂CH₃,        —NHCH₃, —N(CH₃)₂, —N(CH₃)CH₂CH₃ and —N(CH₃)CH₂CH₂CH₃;

-   or R² is a group of formula

wherein

-   -   W is N; the dotted line “a” is a double bond; and R²⁰ is H, OH,        halogen, or Y¹—R^(20a) wherein        -   Y¹ is a bond, O, S, or NR^(20b);        -   R^(20a) is selected from the group consisting of:            (C₁₋₈)alkyl, (C₂₋₈)alkenyl, (C₂₋₈)alkynyl and            (C₃₋₇)cycloalkyl, each of said alkyl, alkenyl, alkynyl and            cycloalkyl being optionally substituted with one, two or            three substituents, each independently selected from:            -   halogen, (C₁₋₆)alkyl, (C₃₋₇)cycloalkyl, Het,                —O—(C₁₋₆)alkyl, —O—(C₃₋₆)cycloalkyl, —NH₂,                —NH(C₁₋₄)alkyl and —N((C₁₋₄)alkyl)₂; and        -   R^(20b) is H, (C₁₋₆)alkyl or (C₃₋₆)cycloalkyl; and    -   R²⁴ is H or (C₁₋₆)alkyl; and    -   R²⁵ is H; and

-   X is O or NH; and

-   R³ is selected from (C₂₋₈)alkyl, (C₃₋₇)cycloalkyl and    (C₃₋₇)cycloalkyl-(C₁₋₃)alkyl-,    -   a) wherein said cycloalkyl and cycloalkyl-alkyl- may be mono-,        di- or tri-substituted with (C₁₋₃)alkyl; and    -   b) wherein said alkyl, cycloalkyl and cycloalkyl-alkyl- may be        mono- or di-substituted with substituents each independently        selected from hydroxy and O—(C₁₋₄)alkyl; and    -   c) wherein each of said alkyl groups may be mono-, di- or        tri-substituted with fluorine or mono-substituted with chlorine        or bromine; and    -   d) wherein in each of said cycloalkyl groups being 5-, 6- or        7-membered, one or two —CH₂-groups not being directly linked to        each other may be replaced by —O— such that the O-atom is linked        to the group X via at least two C-atoms; and        linker D is a 3 to 8 atom saturated or unsaturated alkylene        chain; and        the dotted line “b” is a single bond or a double bond.

More preferred are compounds of formula (I) wherein

-   R¹ is hydroxy or NHSO₂R¹¹, wherein R¹¹ is selected from methyl,    ethyl, 1-methylethyl, propyl, ethenyl, cyclopropyl, cyclobutyl,    cyclopentyl, phenyl,

and —N(CH₃)₂; wherein said phenyl is optionally monosubstituted withhalogen and wherein said cyclopropyl is optionally substituted at the1-position with methyl, ethyl, propyl or butyl, each of said methyl,ethyl, propyl and butyl being optionally further substituted withphenyl, (C₃₋₆)cycloalkyl, (C₂₋₆)alkenyl or (C₁₋₄)alkoxy; and

-   R² is a group of formula

wherein

-   -   R²⁰ is oxo, W is NR²³ wherein R²³ is Me, Et, —CH₂CH═CH₂ or H and        the dotted line “a” is a single bond; or    -   W is N; the dotted line “a” is a double bond; and R²⁰ is H,        (C₁₋₆)alkyl, OH, —O—(C₁₋₆)alkyl, —S—(C₁₋₆)alkyl,        —(CH₂)₀₋₄—CH═CH₂, —(CH₂)₀₋₄—C≡CH, —O—(CH₂)₀₋₄—CH═CH₂,        —O—(CH₂)₀₋₄—C≡CH, —O—(CH₂)₁₋₄—OMe; —O—(CH₂)₁₋₄—N(Me)₂;        —O—(CH₂)₁₋₄-Het; —S—(CH₂)₀₋₄—CH═CH₂, —S—(CH₂)₀₋₄—C≡CH,        —S—(CH₂)₁₋₄—OMe; —S—(CH₂)₁₋₄—N(Me)₂, —S—(CH₂)₁₋₄-Het;        (C₃₋₆)cycloalkyl, —O—(C₃₋₆)cycloalkyl,        O—(C₁₋₆)alkyl-(C₃₋₆)cycloalkyl, —S—(C₃₋₆)cycloalkyl, or        —S—(C₁₋₆)alkyl-(C₃₋₆)cycloalkyl; wherein Het is 5- or 6-membered        monocyclic heteroaryl containing from one to three heteroatoms        each independently selected from N, O and S;        -   each of said (C₁₋₆)alkyl, —O—(C₁₋₆)alkyl, —S—(C₁₋₆)alkyl,            —(CH₂)₀₋₄—CH═CH₂, —(CH₂)₀₋₄—C≡CH, —O—(CH₂)₀₋₄—CH═CH₂,            —O—(CH₂)₀₋₄—C≡CH, —S—(CH₂)₀₋₄—CH═CH₂, —S—(CH₂)₀₋₄—C≡CH,            (C₃₋₆)cycloalkyl, —O—(C₃₋₆)cycloalkyl, and            —S—(C₃₋₆)cycloalkyl being optionally substituted with one,            two or three substituents, each independently selected from            (C₁₋₄)alkyl, —O—(C₁₋₄)alkyl, and halo;    -   or R²⁰ is NR^(20a)R^(20b) wherein R^(20a) is (C₁₋₄)alkyl, and        R^(20b) is H, (C₁₋₄)alkyl or (C₃₋₅)cycloalkyl; and    -   R²¹ is selected from: fluorine, chlorine, bromine, —CH₃,        —CH₂CH₃, —CH₂CH₂CH₃, —OCH₃, —OCH₂CH₃, —OCH₂CH₂CH₃, —SCH₃,        —SCH₂CH₃, —SCH₂CH₂CH₃, —(SO)CH₃, —(SO)CH₂CH₃, —(SO)CH₂CH₂CH₃,        —(SO₂)CH₃, —(SO₂)CH₂CH₃, —(SO₂)CH₂CH₂CH₃,

and —C≡CH; and

-   -   R²² is selected from H, —OCH₃ and —N(CH₃)₂;

-   or R² is a group of formula

wherein

-   -   W is N; the dotted line “a” is a double bond;    -   R²⁰ is Y¹—R^(20a) wherein Y¹ is O and R^(20a) is (C₁₋₈)alkyl;    -   R²⁴ is H or (C₁₋₆)alkyl; and    -   R²⁵ is H; and

-   X is O or NH: and

-   R³ is selected from cyclopropyl, cyclobutyl, cyclopentyl,    cyclohexyl, and a group selected from:

andlinker D is a 3 to 8 atom saturated or unsaturated alkylene chain; andthe dotted line “b” is a single bond or a double bond.

Most preferred are compounds of formula (I) wherein

-   R¹ is hydroxy or NHSO₂R¹¹, wherein R¹¹ is methyl, cyclopropyl,    phenyl,

or —N(CH₃)₂; and

-   R² is a group of formula

wherein

-   -   R²⁰ is oxo, W is NR²³ wherein R²³ is Me, Et, —CH₂CH═CH₂ or H and        the dotted line “a” is a single bond; or    -   W is N, the dotted line “a” is a double bond, and R²⁰ is H,        methyl, ethyl, 1-methylethyl, —C≡CH, O-methyl, O-ethyl, O—        propyl, O—CH(CH₃)₂, O-cyclopentyl, O—CH₂CH₂CF₃, —O—CH═CH₂,        —O—CH₂—CH═CH₂, —O—CH₂—C≡CH, —O—CH₂—C≡CCH₃, —O—CH₂CH₂OMe,        —O—CH₂CH₂N(Me)₂, S-methyl, S-ethyl, S-propyl, S—CH(CH₃)₂,

-   -   -   and

    -   R²¹ is selected from fluorine, chlorine, bromine, —CH₃, —OCH₃,        —SCH₃, —SCH₂CH₃, (SO)CH₃, (SO₂)CH₃, —(SO₂)CH₂CH₃

and —C≡CH; and R²² is H or —OCH₃; or

-   R² is a group of formula

wherein

-   -   W is N; the dotted line “a” is a double bond;    -   R²⁰ is —O—CH₂CH₃;    -   R²⁴ is H or CH₃; and    -   R²⁵ is H; and

-   X is O; and

-   R³ is selected from cyclopentyl,

andlinker D is a 5 carbon atom chain; andthe dotted line “b” is a single bond or a double bond in the Z (cis)configuration.

Specific Examples of Preferred Embodiments

Examples of most preferred compounds according to this invention areeach single compound listed in Tables 1 to 5 below.

Pharmaceutical Composition:

Included within the scope of this invention is a pharmaceuticalcomposition comprising an anti-hepatitis C virally effective amount of acompound of formula (I), or a pharmaceutically acceptable salt or esterthereof, in admixture with at least one pharmaceutically acceptablecarrier medium or auxiliary agent.

According to a further aspect of this embodiment the pharmaceuticalcomposition as defined above further comprises a therapeuticallyeffective amount of at least one other antiviral agent.

According to an alternate embodiment, the pharmaceutical composition ofthis invention may additionally comprise at least one other anti-HCVagent. Examples of anti-HCV agents include, α- (alpha), β- (beta),δ-(delta), γ- (gamma), ω- (omega) or τ- (tau) interferon, pegylatedα-interferon, ribavirin and amantadine.

According to another alternate embodiment, the pharmaceuticalcomposition of this invention may additionally comprise at least oneother inhibitor of HCV NS3 protease.

According to another alternate embodiment, the pharmaceuticalcomposition of this invention may additionally comprise at least oneinhibitor of HCV polymerase.

According to yet another alternate embodiment, the pharmaceuticalcomposition of this invention may additionally comprise at least oneinhibitor of other targets in the HCV life cycle, including but notlimited to, helicase, NS2/3 protease or internal ribosome entry site(IRES).

The pharmaceutical composition of this invention may be administeredorally, parenterally or via an implanted reservoir. Oral administrationor administration by injection is preferred. The pharmaceuticalcomposition of this invention may contain any conventional non-toxicpharmaceutically-acceptable carriers, adjuvants or vehicles. In somecases, the pH of the formulation may be adjusted with pharmaceuticallyacceptable acids, bases or buffers to enhance the stability of theformulated compound or its delivery form. The term parenteral as usedherein includes subcutaneous, intracutaneous, intravenous,intramuscular, intra-articular, intrasynovial, intrasternal,intrathecal, and intralesional injection or infusion techniques.

The pharmaceutical composition may be in the form of a sterileinjectable preparation, for example, as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according totechniques known in the art using suitable dispersing or wetting agents(such as, for example Tween 80) and suspending agents.

The pharmaceutical composition of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, and aqueous suspensions and solutions. Inthe case of tablets for oral use, carriers which are commonly usedinclude lactose and corn starch. Lubricating agents, such as magnesiumstearate, are also typically added. For oral administration in a capsuleform, useful diluents include lactose and dried corn starch. Whenaqueous suspensions are administered orally, the active ingredient iscombined with emulsifying and suspending agents. If desired, certainsweetening and/or flavoring and/or coloring agents may be added.

Other suitable vehicles or carriers for the above noted formulations andcompositions can be found in standard pharmaceutical texts, e.g. in“Remington's Pharmaceutical Sciences”, The Science and Practice ofPharmacy, 19^(th) Ed. Mack Publishing Company, Easton, Pa., (1995).

Dosage levels of between about 0.01 and about 100 mg/kg body weight perday, preferably between about 0.1 and about 50 mg/kg body weight per dayof the protease inhibitor compound described herein are useful in amonotherapy for the prevention and treatment of HCV mediated disease.Typically, the pharmaceutical composition of this invention will beadministered from about 1 to about 5 times per day or alternatively, asa continuous infusion. Such administration can be used as a chronic oracute therapy. The amount of active ingredient that may be combined withthe carrier materials to produce a single dosage form will varydepending upon the host treated and the particular mode ofadministration. A typical preparation will contain from about 5% toabout 95% active compound (w/w). Preferably, such preparations containfrom about 20% to about 80% active compound.

As the skilled artisan will appreciate, lower or higher doses than thoserecited above may be required. Specific dosage and treatment regimensfor any particular patient will depend upon a variety of factors,including the activity of the specific compound employed, the age, bodyweight, general health status, sex, diet, time of administration, rateof excretion, drug combination, the severity and course of theinfection, the patient's disposition to the infection and the judgmentof the treating physician. Generally, treatment is initiated with smalldosages substantially less than the optimum dose of the peptide.Thereafter, the dosage is increased by small increments until theoptimum effect under the circumstances is reached. In general, thecompound is most desirably administered at a concentration level thatwill generally afford antivirally effective results without causing anyharmful or deleterious side effects.

When the composition of this invention comprises a combination of acompound of formula (I) and one or more additional therapeutic orprophylactic agent, both the compound and the additional agent should bepresent at dosage levels of between about 10 to 100%, and morepreferably between about 10 and 80% of the dosage normally administeredin a monotherapy regimen.

When these compounds, including their pharmaceutically acceptable saltsand esters thereof, are formulated together with a pharmaceuticallyacceptable carrier, the resulting composition may be administered invivo to mammals, such as man, to inhibit HCV NS3 protease or to treat orprevent HCV virus infection. Such treatment may also be achieved using acompound of this invention in combination with another antiviral agent.Preferred other antiviral agents are described within the Definitionssection and the section of preferred pharmaceutical compositionsaccording to this invention and include, but are not limited to: α-(alpha), β- (beta), δ-(delta), ω- (omega), γ- (gamma) or τ-(tau)-interferon, ribavirin, amantadine; other inhibitors of HCV NS3protease; inhibitors of HCV polymerase; inhibitors of other targets inthe HCV life cycle, which include but are not limited to, helicase,NS2/3 protease, or internal ribosome entry site (IRES); or combinationsthereof. The additional agents may be combined with compounds of thisinvention to create a single dosage form. Alternatively these additionalagents may be separately administered to a mammal as part of a multipledosage form.

Accordingly, another embodiment of this invention provides a method ofinhibiting HCV NS3 protease activity in a mammal by administering acompound of the formula (I), including a pharmaceutically acceptablesalt or ester thereof.

In a preferred embodiment, this method is useful in decreasing the NS3protease activity of the hepatitis C virus infecting a mammal.

Still another embodiment of this invention provides the use of acompound of formula (I),

or a pharmaceutically acceptable salt or ester thereof, to inhibit HCVNS3 protease activity.

As discussed above, combination therapy is contemplated wherein acompound of formula (I), or a pharmaceutically acceptable salt or esterthereof, is co-administered with at least one additional antiviralagent. Preferred antiviral agents are described hereinbefore andexamples of such agents are provided in the Definitions section. Theseadditional agents may be combined with the compounds of this inventionto create a single pharmaceutical dosage form. Alternatively theseadditional agents may be separately administered to the patient as partof a multiple dosage form, for example, using a kit. Such additionalagents may be administered to the patient prior to, concurrently with,or following the administration of a compound of formula (I), or apharmaceutically acceptable salt or ester thereof.

A compound of formula (I), or a pharmaceutically acceptable salt orester thereof, set forth herein may also be used as a laboratoryreagent. Furthermore a compound of this invention, including apharmaceutically acceptable salt or ester thereof, may also be used totreat or prevent viral contamination of materials and therefore reducethe risk of viral infection of laboratory or medical personnel orpatients who come in contact with such materials (e.g. blood, tissue,surgical instruments and garments, laboratory instruments and garments,and blood collection apparatuses and materials).

A compound of formula (I), including a pharmaceutically acceptable saltor ester thereof, set forth herein may also be used as a researchreagent. A compound of formula (I), including a pharmaceuticallyacceptable salt or ester thereof, may also be used as positive controlto validate surrogate cell-based assays or in vitro or in vivo viralreplication assays.

Methodology

In general, the compound of formula (I) and intermediates therefore areprepared by known methods using reaction conditions which are known tobe suitable for the reactants. Several such methods are disclosed in WO00/09543, WO 00/09558, WO 00/59929 and in co-pending application Ser.No. 10/945,518, all of which are incorporated herein by reference.

Particularly, the synthesis of the P3 fragment ((2S)—N-protected-aminonon-8-enoic acid) and the P1 fragment ((1R,2S)1-amino-2-ethenylcyclopropylcarboxylic acid) have been described indetail in WO 00/59929.

I. General Multi-Step Synthetic Method

In general, the present invention is directed to compounds of formula(I) which can be prepared by a general multi-step synthetic method.Specifically, compounds of the following formula (Ib) are prepared bythe following process:

wherein P1, R¹, R², and D are as defined herein and wherein R^(3a) isdefined as —NHC(═O)—X—R³, wherein X and R³ are as defined herein,said process comprising the following steps:(i) reacting a compound of formula (II):

or a salt thereof, with a compound of formula (III):

(ii) reacting the resulting compound of formula (IV) obtained in step(i):

with an aminocyclopropane compound of formula (V)

(iii) reacting the resulting compound of formula (VI) obtained in step(ii):

with a compound of formula (VII):

LG-SO₂—R¹²  (VII)

wherein LG represents a suitable leaving group and R¹² is selected fromp-tolyl, p-bromophenyl, p-nitrophenyl, methyl, trifluoromethyl,perfluorobutyl and 2,2,2-trifluoroethyl;iv) cyclizing of the resulting diene compound of formula (VIII) obtainedin step (iii):

in the presence of a ruthenium catalyst; and(v) reacting the resulting compound of formula (IX) obtained in step(iv):

with a compound of formula (X):

-   -   wherein R² is selected from

-   -   wherein W, R²⁰, R²¹, R²², R²⁴, R²⁵ and the dotted line a are as        defined herein;        to obtain a compound of formula (Ib):

and when R¹ is a carboxylic acid ester group in the resulting compoundof formula (Ib), optionally subjecting the compound of formula (Ib) tohydrolysis conditions to obtain a compound of formula (I) wherein R¹ isa carboxylic acid group.

II. Sulfonamides and Sulfamides

Compounds of formula (I) wherein R¹ is NHSO₂R¹¹ as defined herein areprepared by coupling the corresponding acid of formula (I) (i.e. R¹ ishydroxy) with an appropriate sulfonamide or sulfamide of formulaR¹¹—SO₂NH₂ in the presence of a coupling agent under standardconditions. Although several commonly used coupling agents can beemployed, TBTU and HATU have been found to be practical. Thesulfonamides are available commercially or can be prepared by knownmethods.

III. Alternative Methodology

The following scheme provides an alternative process using known methodsfor preparing a key intermediate of formula 1h from acyclicintermediates:

Scheme I:

Steps A, C, D: Briefly, the P1, P2, and P3 moieties can be linked bywell known peptide coupling techniques generally disclosed in WO00/09543 & WO 00/09558.

Step B: This step involves the inversion of configuration of the4-hydroxy substituent. There are several ways in which this can beaccomplished as will be recognized by persons skilled in the art. Oneexample of a convenient method is the well known Mitsunobu reaction(Mitsunobu Synthesis 1981, Jan. 1-28; Rano et al. Tet. Lett. 1994, 36,3779-3792; Krchnak et al. Tet. Lett. 1995, 36, 6193-6196).

Step E: The formation of the macrocycle can be carried out via an olefinmetathesis using a Ru-based catalyst such as the one reported by Miller,S. J.; Blackwell, H. E.; Grubbs, R. H. J. Am. Chem. Soc. 1996, 118,9606-9614 (a); Kingsbury, J. S.; Harrity, J. P. A.; Bonitatebus, P. J.;Hoveyda, A.H. J. Am. Chem. Soc. 1999, 121, 791-799 (b) and Huang, J.;Stevens, E. D.; Nolan, S. P.; Petersen, J. L.; J. Am. Chem. Soc. 1999,121, 2674-2678 (c) or as described in WO 00/59929. It will also berecognized that catalysts containing other transition metals such as Mocan be used for this reaction.

Step F: Conversion of the hydroxyl group of the proline to a suitableleaving group (i.e. brosylate) was carried out by reacting the free OHwith the corresponding halo-derivative (i.e. 4-bromobenzenesulfonylchloride).

Subsequent conversion of the key intermediate of formula 1h to thecompounds of formula (I) of this invention is disclosed in detail in theexamples hereinafter.

IV. Introduction of the R² Moiety to Form Compounds of General Formula(Ic):

The general process comprises reacting a macrocyclic compound of formula(IXa or 1h) with a compound of formula (X):

Compounds of formula (IXa) and (X) are mixed in a polar non-proticorganic solvent (such as THF, dioxane, dichloromethane, chloroform,N-methylpyrrolidone, dimethyl sulfoxide, dimethylformamide, acetone, ormethylisobutylketone) in the presence of an inorganic or organic base(such as cesium carbonate, or DBU) at 40° C. to 100° C. until completionof reaction. Aqueous workup followed by crystallization from a suitablesolvent such as ethyl acetate-heptane or ethyl acetate/methylcyclohexaneprovides the compounds of formula (Ic).

VI. Synthesis of P2 Substituents:

The compounds of formula (X) used as starting material may besynthesized from commercially available materials using the techniquesdescribed in the literature.

General Protocol for the Preparation of 2-alkoxy Substituted4-hydroxyquinolines (2a):

Compounds of formula X wherein R² is a group of formula

where R²⁰ is an alkoxy group, can be prepared according to the followingscheme 2:

Briefly, following the known Pinner synthesis, a suitably functionalizedcyanoester is condensed with the corresponding alcohol using a fullysaturated HCl/Et₂O solution [Neilson, in Patai, “The Chemistry ofAmidines and Imidates.” pp. 385-489, Wiley, N.Y., 1975.]. The resultingimidate salt is then subsequently condensed with an appropriatelysubstituted aniline to form the aniline derived imidate. Thermalcyclization affords the corresponding 2-alkoxy substituted4-hydroxyquinolines 2a.

For example, when R^(20a)=Et in the above scheme, ethyl cyanoacetate andethanol are used as reagents.

For R^(20a)=Me in the above scheme, methyl cyanoacetate and methanol areused as reagents.

Compounds of formula X wherein R² is a group of formula

and R²⁰ is an alkoxy group can also be prepared according to scheme 2,by using an appropriately substituted aminothiophene in place of theaniline in step B above.General Protocol for the Preparation of 2-alkyl substituted4-hydroxyquinolines (2b):

Compounds of formula X wherein R² is a group of formula

where R²⁰ is an alkyl group, can be prepared according to the followingscheme 3:

Briefly, appropriately substituted β-ketoesters are condensed withsubstituted anilines and subsequently thermally cyclized to afford thecorresponding 2-alkyl substituted hydroxyquinolines. For example, whenthe initial condensation reaction with the aniline (step A) is performedwith the corresponding methyl ketone (R²⁰═CH₃), a methyl group isincorporated in the 2-position of the resulting 4-hydroxyquinoline.

General Protocol for the Preparation of 2-thioalkyl Substituted4-hydroxyquinolines (2c):

In general, the various 2-thioalkyl analogs were prepared as shown inthe following scheme 4.

Briefly, condensation of diethyl malonate under basic conditions with asuitably functionalized isothiocyanate produces the malonate adduct as asalt. Treatment of the salt with an alkylating reagent (e.g. EtI: ethyliodide) produces a mixture of S- and N-alkylated products. Thermalcyclization of this mixture gives the 3-ethyl carboxylate which issaponified and decarboxylated to produce the desired 2-thioalkylsubstituted hydroxyquinolines. For example, utilization of EtI in thealkylation step results in the formation of the 2-thioethyl analog(R^(20a)=Et).

Alternative Protocol for the Preparation of 2-alkoxy and 2-alkynylSubstituted 4-hydroxyquinolines (2d):

Compounds of formula X wherein R² is a group of formula

wherein R²⁰ is a substituted or unsubstituted alkoxy group or an alkynylgroup may be prepared according to the following Scheme 5:

Briefly, a compound of formula 2c (Scheme 4), wherein R^(20a) is a smallalkyl group such as ethyl or propyl, is protected at the hydroxy groupas, for example, a p-methoxybenzyl (PMB) ether. The protected quinolineis oxidized, using reagents well known in the art, to give a sulfonewhich is then treated with a suitable nucleophile to introduce the R²⁰group. Examples of suitable nucleophiles include, but are not limitedto, appropriately substituted and/or protected alkoxide anions andacetylide anions. Deprotection of the PMB ether, and of the R²⁰substituent if necessary, then gives the desired 4-hydroxyquinolinecompounds 2d.

The corresponding anilines are commercially available or may requiresome well known chemical transformations. For example if the nitroanalog is commercially available, it can be converted to thecorresponding amine by using any of several reducing agents well knownto one skilled in the art. Also, if the carboxylic acid is commerciallyavailable, transformation into the corresponding amine is possible via aCurtius rearrangement.

Further details of the invention are illustrated in the followingexamples which are understood to be non-limiting with respect to theappended claims. Other specific ways of synthesis or resolution of thecompounds of this invention can be found in WO 99/07733, WO 00/09543; WO00/09558 & WO 00/59929 and in co-pending application Ser. No.10/945,518, all of which are hereby incorporated by reference.

EXAMPLES

Temperatures are given in degrees Celsius. Solution percentages expressa weight to volume relationship, and solution ratios express a volume tovolume relationship, unless stated otherwise. Nuclear magnetic resonance(NMR) spectra were recorded on a Bruker 400 MHz spectrometer; thechemical shifts (δ) are reported in parts per million and are referencedto the internal deuterated solvent unless otherwise indicated. The NMRspectra of all final compounds (inhibitors) was recorded in DMSO-d₆.Flash column chromatography was carried out on silica gel (SiO₂)according to Still's flash chromatography technique (W. C. Still et al.,J. Org. Chem., 1978, 43, 2923). Analytical HPLC was carried out understandard conditions using a Combiscreen ODS-AQ C18 reverse phase column,YMC, 50×4.6 mm i.d., 5 μM, 120 A at 220 nM, elution with a lineargradient as described in the following table (Solvent A is 0.06% TFA inH₂O; solvent B is 0.06% TFA in CH₃CN):

Time (min) Flow (mL/min) Solvent A (%) Solvent B (%) 0 3.0 95 5 0.5 3.095 5 6.0 3.0 50 50 10.5 3.5 0 100

Abbreviations used in the examples include Boc: tert-butyloxycarbonyl[Me₃COC(O)]; BSA: bovine serum albumin; Brs: brosyl(p-bromobenzenesulfonyl); CDI: N,N′-carbonyldiimidazole; CHAPS:3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate; DABCO:1,4-diazabicyclo[2.2.2]octane; DBU: 1,8-diazabicyclo[5.4.0]undec-7-ene;DCM: dichloromethane (CH₂Cl₂; methylene chloride); DCE: dichloroethane;DCHA: dicyclohexylamine; DEAD: diethylazodicarboxylate; DIAD:diisopropylazodicarboxylate; DIPEA: diisopropylethylamine; DMAP:dimethylaminopyridine; DMF: N,N-dimethylformamide; DMSO:dimethylsulfoxide; EDCI:1-ethyl-3-[3-(dimethylamino)propyl]-carbodiimide hydrochloride;(S,S)-Et-DUPHOS Rh (COD)OTf: (+)-1,2-bis(2S,5S)-2,5-diethylphospholano)benzene (cyclooctadiene) rhodinium (1) trifluoromethanesulfonate; Et:ethyl; EtOH: ethanol; EtOAc: ethyl acetate; ESMS: electrospray massspectrometry; FAB: Fast Atom Bombardment; HATU:O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate; HPLC: high performance liquid chromatography; MS:mass spectrometry; MALDI-TOF: Matrix Assisted Laser DisorptionIonization-Time of Flight; MCH: methylcyclohexane; Me: methyl; MeOH:methanol; MIBK: methyl isobutyl ketone; NMP: N-methylpyrrolidinone; PMB:para-methoxy benzyl; Pr: propyl; R.T.: room temperature (18° C.-22° C.);TBTU: 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumtetrafluoroborate; TFA: trifluoroacetic acid; THF: tetrahydrofuran; THP:tetrakishydroxymethyl phosphonium chloride; TLC: thin layerchromatography; Tris/HCl: tris(hydroxymethyl)aminomethane hydrochloride;SEH: sodium 2-ethylhexanoate; PTSA: para-toluenesulfonic acid.

Example 1 Synthesis of INRF12 Brosylate Intermediate Step 1:Introduction of the Boc-Protecting Group; Synthesis of INRF2

The amino-protection was done with the Boc-protecting-group. INRF 1(trans-4-hydroxy L-proline) (249.8 g, 1.905 mol) was dissolved in water(375 mL) and 45% sodium hydroxide solution (203 g, 2.286 mol). To ensuregood phase transfer, tert-butanol (106 g) was added. In an alternativeprocedure, acetone was used instead of THF/tert-butanol. The reactionmixture was heated to 50° C. and the anhydride Boc₂O (424 g, 1.943 mol),dissolved in THF (425 mL), or acetone, was slowly added. The reaction isexothermic and generates gas (CO₂) as the Boc₂O was added. If thereaction does not proceed as desired, catalytic amounts of DMAP (2.3 g,19 mmol) can be added. After the addition of the Boc₂O, the reactionmixture was kept 0.5-1 h at 50° C., and the THF was removed by partialdistillation. The pH of the remaining solution was adjusted to about pH3 with concentrated HCl (204 g, 2.076 mol) and the product was thenextracted with MIBK (1 liter) and again with MIBK (375 mL). The organiclayer was heated and some of the solvent was distilled off to removetraces of water. The product was crystallized from this solution byadding MCH (1.25 L), isolated by filtration, washed twice with MCH (375mL) and dried overnight at 40° C.

Yield: 77-78%, colorless crystals, F_(p)=126-128° C.

Step 2: Formation of the Lactone; Synthesis of PDIG0016

INRF 2 (416.3 g, 1.8 mol) is dissolved in THF (2.08 L) and cooled withice to a temperature from about −5 to −10° C. Mesylchloride (392 g, 3.4mol) and N-methylpyrrolidine (429 g, 5 mol) is added and the mixturestirred for about 1½ at about −5° C. The mixture is washed with waterand heated to reflux. Dioxane (2.08 L) is added and the THF is distilledoff. After cooling down to room temperature, DIPEA (233 g, 1.8 mol) isadded and the mixture is heated to reflux. After 1 h, part of thesolvent (830 mL) is distilled off. The remaining solution is cooled toambient temperature, a KHSO₄-solution (14.4 g in 2.08 l water) is addedand the solution is allowed to cool down to room temperature. Theresulting crystals (PDIG0016) are isolated by filtration, washed withwater and dried overnight at 45° C.

Yield: 78-82%, colorless needles, F_(p)=111° C.

Step 3: Deprotection of the Lactone; Synthesis of PDIG0017MS

The lactone PDIG0016 (267 g, 1.25 mol) is dissolved inmethyl-isobutylketone (1467 mL). The suspension is heated up to 50° C.until the lactone is completely dissolved and a part of the solvent (130mL) is distilled off to remove traces of water. Methanesulfonic acid(240 g, 2.5 mol) is added slowly to the reaction mixture. During theaddition, gas is evolved (CO₂, isobutene). The reaction mixture isallowed to cool to room temperature and the resulting crystals areisolated by filtration, washed twice with acetone (each 400 mL) anddried overnight at 40° C.

Yield: 93-98%, colorless crystals, 208-210° C.

Step 4: Coupling with INRF 15; Synthesis of the Dipeptide PDIG0027

First, INRF15-DCHA has to be released. Therefore, INRF15-DCHA (61.4 g,132 mmol) is dissolved in toluene (160 mL) and the resulting solution iswashed with diluted sulfuric acid (5.3 g in 80 mL water) and water (80mL). After phase separation, the solution is treated with charcoal andfiltered and the resulting solution stored at room temperature.

The deprotected lactone PDIG0017MS (24.9 g, 119 mmol) and EDCI.HCl (26.8g, 140 mmol) are suspended in dichloromethane (140 mL) and cooled toroom temperature. The suspension is treated with the INRF15-solutionpreviously generated. To this suspension, di-isopropylethylamine(Hünigs-Base, 16.3 g, 130 mmol) is slowly added while the reaction iskept under nitrogen at temperatures below 20° C. The suspension isfiltered, and the resulting solution is washed with water (80 mL),diluted acetic acid (1.3 g in 80 mL water), 5% sodium bicarbonatesolution (80 mL) and again with water (80 mL). After phase separation,dichloromethane is distilled off under reduced pressure. The resultingsolution can directly be used for the next step. Otherwise, the productcan be isolated by crystallization from MCH.

Yield: 95% (GC), yellowish solution, F_(p)=58-60° C.

Step 5: Synthesis of INRF 16-OH

A mixture of PDIG0027 (10.0 g, 23.7 mmol, 1.0 eq.), INRF3 (7.6 g, 24.2mmol, 1.02 eq.) and sodium 2-ethylhexanoate (SEH) (5.9 g, 35.6 mmol, 1.5eq.) in water (43 mL) and toluene (12 mL) is stirred at 80° C. for 2 h.For work-up, toluene (75 mL) is added at 80° C. After stirring andseparation of the aqueous layer, the organic layer is washed with 1MNa₂CO₃ (3×30 mL), 0.5M HCl (30 mL) and water (2×30 mL). The solvent isremoved under vacuum.

Yield of INRF16—OH: 11.7 g, 22.5 mmol, 95%; purity: >95% (peak-areaHPLC) as a slightly yellow oil.

Step 6. Brosylation of INRF16—OH; Synthesis of INRF16-Brs

To a mixture of INRF16-OH (10.7 g, 18.5 mmol, 1.0 eq.) and DABCO (3.3 g,29.7 mmol, 1.6 eq.) and toluene (23 mL) a solution of4-bromobenzenesulfonyl chloride (brosyl chloride, 6.6 g, 26.0 mmol, 1.4eq.) in toluene (15 mL) is added slowly at room temperature. The mixtureis stirred for 2 h. For work-up the organic layer is washed with 1MNa₂CO₃ (2×21 mL), diluted with THF (21 mL) and washed with 0.5M HCl (21mL) and water (2×21 mL). The solvent is removed under vacuum.

Yield of INRF16-Brs: 12.3 g, 16.7 mmol, 90%; purity: >95% (peak-areaHPLC) as a slightly orange oil. A charcoal treatment of the crudeproduct is possible.

Step 7: Metathesis of INRF16Brs to INRF12Brs

Preparation of the THP-solution (for an experiment with 35.4 gINRF16Brs): 23.5 g of tetrakis(hydroxymethyl)phosphonium chloride (80%in water, 98.7 mmol) is dissolved in isopropanol (35 mL) under anitrogen atmosphere. Then 12.1 g (98.7 mmol) of a 45% KOH solution isadded within 5 min while the solution is cooled (temperature 20-25° C.).After stirring the suspension for another 30 min under nitrogen, themixture is filtered and the inorganic residue is washed with 20 mL ofdegassed isopropanol. The combined isopropanol solution is stored undera nitrogen atmosphere until use.

Metathesis Reaction:

In a reaction flask 3500 mL of toluene is degassed by bubbling nitrogenthrough the toluene. 35.2 g (47.7 mmol) of INRF16Brs are dissolved in 70mL of degassed toluene and added into the reaction flask. The solutionis heated up to 80° C. and 3 mol % of Hoveyda's catalyst is added undernitrogen in four portions over a period of 3 hours. After stirring for afurther 60 min at the same temperature the conversion is checked byHPLC. In the case that the conversion is below 95%, additional Hoveyda'scatalyst is added and the mixture is stirred until the conversionis >95% (during the reaction a slight stream of nitrogen is bubbledthrough the reaction mixture).

After cooling to 50° C. the THP solution is added to the reactionmixture. After stirring for 8.5 h at 50° C. the mixture is cooled toroom temperature and extracted twice with 188 mL of degassed water, 188mL of 0.5 M HCl, 188 mL of 0.5 M NaHCO₃ solution, and 188 mL of water.

Approximately 2800 mL of toluene are distilled off at 50° C. underpartial reduced pressure and the remaining solution is treated at 50° C.with 6.8 g of activated charcoal which is then removed by filtration.

The remaining liquid filtrate (approx. 130 mL) is added over a period of1 hour to 1.5 liters of pre-cooled MCH (5° C.). After stirring for afurther 30 min at 5° C. the precipitate is filtered and washed with 100mL of MCH (several portions). The white solid is dried in vacuo at 25°C.

Yield (by weight): 38 g of an almost white powder.

Example 1A Synthesis of Brosylate Intermediate with a Saturated Linkerfor the Synthesis of Compounds of Table 2

Step A: To brosylate INRF12-Brs (1.0 g, 1.41 mmol, 1 eq.) dissolved inEtOAc (30 mL) was added 5% Rh/Al (300 mg, 30% w/w). The atmosphere wassaturated with H₂ gas and stirred at RT for 10 h until completeconversion by HPLC analysis. The suspension was filtered through Celiteto remove the catalyst and the solvent removed in vacuo to afford awhite solid. Purification (SiO₂, (1:1) EtOAc/hexanes) afforded afterconcentration the saturated macrocycle 1A1 (467 mg, 47%) as a whitesolid.

Example 2A Synthesis of 1-methyl-2-methoxy aniline (2A2)

To a solution of 2-methyl-3-nitro anisole (2A1) (5.1 g; 30.33 mmol;requires ˜30 min. to dissolve) in absolute ethanol (85 mL) was added 10%Pd/C catalyst (500 mg). The solution was hydrogenated under a hydrogenfilled balloon at atmospheric pressure and room temperature for 19 h.The reaction mixture was filtered through a Celite pad, rinsed andevaporated to dryness to obtain the compound 2A2 as a deep mauve oil(4.1 g 29.81 mmol; 98% yield).

MS 137 (MH)+. Reverse Phase HPLC Homogeneity @ 220 nm (0.06% TFA;CH₃CN:H₂0):99%.

Example 2B Synthesis of 2-bromo-3-methoxy aniline (2B4)

Step A:

2-Amino-3-nitrophenol 2B1 (5 g; 32.4 mmol) was dissolved in H₂O (29.5mL) and 1,4-dioxane (14.7 mL). The mixture was heated to reflux andhydrobromic acid (48%; 16.7 mL; 147 mmol) was added dropwise over aperiod of 20 min. Upon completion of the addition, the reflux wasmaintained an additional 15 min. The reaction was cooled to 0° C. (icebath), and sodium nitrite (2.23 g; 32.3 mmol) in H₂O (20 mL) was addedover a period of 30 min. The stirring was continued for 15 min. at 0°C., the mixture transferred to a jacketed dropping funnel (0° C.) andadded dropwise to a stirred mixture of Cu(1)Br (5.34 g; 37.2 mmol) inH₂O (29.5 mL) and HBr (48%; 16.7 mL; 147 mmol) at 0° C. The reaction wasstirred for 15 min. at 0° C., warmed to 60° C., stirred for anadditional 15 min., cooled to room temperature, and left to stirovernight. The reaction mixture was transferred to a separatory funneland extracted with ether (3×150 mL). The organic layers were combined,washed with brine (1×), dried (Na₂SO₄), filtered and concentrated toafford the crude product (7.99 g) as a red-brown oil. The crude materialwas purified by flash column chromatography (1:25 ultra pure silica gel,230-400 mesh, 40-60 mm, 60 angstroms; CH₂Cl₂ as the solvent) to affordpure 2-bromo-3-nitro-phenol 2B2 (45%; 3.16 g) as an orange-brown solid.

MS 217.8 (MH)⁻. Homogeneity by HPLC (TFA) @ 220 nm: 97%.

Step B:

The nitrophenol starting material 2B2 (3.1 g; 14.2 mmol) was dissolvedin DMF (20 mL) and to the solution was added ground cesium carbonate(5.58 g; 17.1 mmol) followed by MeI (2.6 mL; 42.5 mmol). The mixture wasstirred at room temperature overnight. The DMF was evaporated, theresidue taken up in ether (1×200 mL), washed with water (1×200 mL),brine (4×100 mL), dried (MgSO₄), filtered and evaporated to afford thecrude 2-bromo-3-nitroanisole 2B3 (94%; 3.1 g) as an orange solid.

MS 234 (M+2H)⁺; Homogeneity by HPLC (TFA) @ 220 nm: 98%

Step C:

2-Bromo-3-nitroanisole 2B3 (1.00 g; 4.31 mmol) was dissolved in glacialacetic acid (11.0 mL)/ethanol (11.0 mL) and to the solution was addediron powder (0.98 g; 17.5 mmol). The mixture was stirred at reflux for3.5 h and worked up. The reaction mixture was diluted with water (35mL), neutralized with solid Na₂CO₃ and the product extracted with CH₂Cl₂(3×50 mL). The extracts were dried (Na₂SO₄), filtered and concentratedin vacuo to afford the crude product, 2-bromo-3 methoxyaniline 2B4 (91%;0.79 g) as a pale yellow oil.

MS 201.8 (MH)⁺; Homogeneity by HPLC (TFA) @ 220 nm: 95%

Example 2C Synthesis of 2-chloro-3-methoxy aniline (2C3)

Step A:

2-Amino-3-nitrophenol 2B1 (5 g; 32.4 mmol) was dissolved in concentratedHCl (75 mL) and 1,4-dioxane (14.7 mL). The mixture was heated to 70° C.until most of the solids were in solution. The reaction mixture wascooled to 0° C. (ice bath), and sodium nitrite (2.23 g; 32.3 mmol) inH₂O (5.4 mL) was added over a period of 3 hours to the brown solution.The temperature was maintained below 10° C. during the addition and thestirring was continued for an additional 15 min. at 0° C. This diazoniumintermediate was poured into a solution of Cu(I)Cl (3.8 g; 38.9 mmol) inH₂O (18.5 mL) and conc. HCl (18.5 mL) at 0° C. The reaction was stirredfor 15 min. at 0° C., warmed to 60° C., and stirred for an additional 15min. The reaction mixture was then brought to room temperature, and leftto stir overnight. The reaction mixture was transferred to a separatoryfunnel and extracted with ether (3×150 mL). The organic layers werecombined, washed with brine (1×), dried (Na₂SO₄), filtered andconcentrated to afford the crude product (5.83 g) as a red-brown oil.The crude material was purified by flash column chromatography (1:25ultra pure silica gel, 230-400 mesh, 40-60 mm, 60 angstroms; 3:1hexane/EtOAc as the solvent) to afford pure 2-chloro-3-nitrophenol 2C1(48%; 2.7 g) as an orange solid.

MS 171.8 (MH)⁻: Homogeneity by HPLC (TFA) @ 220 nm: 96%.

Relevant literature for the Sandmeyer Reaction: J. Med. Chem., 1982,25(4), 446-451.

Step B:

The nitrophenol starting material 2C1 (1.3 g; 7.49 mmol) was dissolvedin DMF (10 mL) and to this solution was added ground cesium carbonate(2.92 g; 8.96 mmol), followed by MeI (1.4 mL; 22.5 mmol). The mixturewas stirred at room temperature overnight. The DMF was evaporated invacuo and the residue taken up in ether (150 mL), washed with water (150mL), brine (4×100 mL), and then dried over (MgSO₄). The organic phasewas filtered and evaporated to afford the crude 2-chloro-3-nitroanisole2C2 (98%; 1.38 g) as an orange solid.

Homogeneity by HPLC (TFA) @ 220 nm: 93%.

Step C:

2-Chloro-3-nitroanisole 2C2 (1.38 g; 7.36 mmol) was dissolved in amixture of glacial acetic acid (19 mL)/ethanol (19 mL). To this solutionwas added iron powder (1.64 g; 29.4 mmol). The mixture was stirred atreflux for 3.5 h and worked up. The reaction mixture was diluted withwater (70 mL), neutralized with solid Na₂CO₃ and the product extractedwith CH₂Cl₂(3×150 mL). The extracts were combined and washed with sat.brine and then dried over (Na₂SO₄), filtered and concentrated in vacuoto afford the crude product, 2-chloro-3-methoxyaniline 2C3 (100%; 1.2 g)as a yellow oil. This material was used as such in the following steps.

MS 157.9 (MH)⁺; Homogeneity by HPLC (TFA) @ 220 nm: 86%.

Example 3A Preparation of 2-ethoxy-4-hydroxy-8-chloroquinoline (3A5)

Step A: To ethyl cyanoacetate 3A1 (23 g, 0.203 mol) was added absoluteethanol (10 g, 12.7 mL, 0.22 mol) in diethyl ether (20 mL). The solutionwas cooled to 0° C. in an ice bath before being treated with HCl gas(bubbled through solution for 12 minutes resulted in an increase inweight of 12 g (˜0.33 mol)).

This solution was stirred at 0° C. for 6 h and then was allowed to warmto R.T. and was stirred for 16 h. The resultant solid was broken up andwashed several times with ether and then placed in vacuo for severalhours. The imidate salt 3A2 was obtained as a white solid (36.4 g, 92%)and was stored under a nitrogen atmosphere. The ¹H NMR is consistentwith the desired product.

Step B: The imidate salt 3A2 (1.47 g, 7.5 mmol, 1 eq.) was combined with2-chloroaniline 3A3 (0.96 g, 7.50 mmol, 1 eq.) in ethanol (15 mL) underan N₂ atmosphere. The reaction mixture was stirred at R.T. (16 h) andmonitored by HPLC. The reaction mixture was concentrated and thenpurified directly over silica gel (eluent: 10% EtOAc/hexanes) to affordthe condensation product 3A4 as a clear oil (1.73 g, 86%). MSelectrospray: (MH)+; 270 and (M−H)−; 268. TLC (UV) Rf=0.50 (10%EtOAc/hexane).

Step C: The condensation product 3A4 (1.73 g, 6.41 mmol) was dissolvedin diphenyl ether (10 mL) and placed in a sand bath (300° C.). Theinternal temperature was monitored and maintained between 240-250° C.for 8 minutes. The mixture was cooled and then directly loaded on asilica gel column and eluted first with hexanes, then with 30%EtOAc/hexanes and finally 50% EtOAc/hexanes. The product wasconcentrated and dried in vacuo to give the corresponding4-hydroxyquinoline derivative 3A5 as a beige crystalline solid (0.76 g,53%). MS electrospray: (M+H)⁺; 224 and (M−H)⁻; 222.

Example 3B Preparation of 4-hydroxy-8-chloroquinoline (3B3)

Step A: To 2-chloroaniline 3A3 (1.6 mL, 15.2 mmol, 1 eq) dissolved inanhydrous acetonitrile (50 mL) at R.T. was added Meldrum's acid 3B1(2.41 g, 16.73 mmol, 1.1 eq), followed by trimethyl orthoformate (2.0mL, 18.25 mmol, 1.2 eq). The resulting mixture was heated to reflux (95°C.) for 2 h and monitored by analytical HPLC until complete. Theresulting solution was cooled to R.T. and evaporated to dryness toafford a beige solid that was recrystallized from boiling MeOH. Afterdrying in vacuo adduct 3B2 was obtained as a bright yellow solid (2.29g, 53%).

Step B: In a pre-heated sand bath (300-350° C.), diphenyl ether (6 mL)was heated until the internal temperature reached 220° C. Adduct 3B2(981 mg, 3.48 mmol) was added portionwise over ca. 4 min period (gasevolution) to the heated solvent. The temperature (220° C.) wasmaintained for another 5 min after which the solution was allowed tocool. Upon cooling, the product precipitated out of solution and wasfiltered and washed with diethyl ether. After drying in vacuo (16 h),product 3B3 was obtained as a beige solid (417 mg, 67%). MS: (M+H)+;180.

Example 3C Preparation of 8-chloro-4-hydroxy-2-methylquinoline (3C3)

Step A: To a solution of ethyl acetoacetate 3C1 (1.21 mL, 9.51 mmol; 1eq) in benzene (20 mL) was added 2-chloroaniline 3A3 (1.0 mL; 9.51 mmol;1 eq) followed by catalytic PTSA (13 mg). The reaction flask wasequipped with a Dean-Stark apparatus and heated to reflux for 2 hours.The solvent was removed and the residue purified by columnchromatography using silica gel (eluent: 10% EtOAc/hexanes; R_(f)=0.48)to give compound 3C2 (1.46 g, 64%) as a clear oil. MS: (M+H)+; 240, HPLChomogeneity=99.5%.

Step B: In a pre-heated sand bath (300-350° C.), compound 3C2 (730 mg,3.0 mmol) in diphenyl ether (8 mL) was heated until the internaltemperature reached 220° C. and that temperature was maintained for 7minutes after which the solution was allowed to cool. Upon cooling, abeige solid precipitated out and was filtered and washed with diethylether. After drying, the desired quinoline 3C3 was obtained as a beigesolid (452 mg, 77%). MS: (M+H)+; 194, HPLC homogeneity=99%.

Example 3D Preparation of 2-thioethyl-8-chloro-4-hydroxyquinoline (3D7)

Step A: To THF (30 mL) was added sodium hydride (60% in oil, 920 mg, 23mmol, 1.2 eq) before being cooled to 0° C. Diethyl malonate (2.91 mL,19.15 mmol, 1.0 eq) was then added dropwise (gas evolution) and thissolution was allowed to warm to R.T. and was stirred for 1 h. Thismixture was cooled down to 0° C. before the addition of 2-chlorophenylisothiocyanate 3D1 (2.5 mL, 19.15 mmol, 1.0 eq). The resulting mixturewas again allowed to warm to R.T. for 3 h until the SM was consumed. Theorange solution was concentrated down and dried in vacuo to afford thesodium salt adduct 3D2 (6.73 g, 100%) as an orange crystalline solid.This material can be used as is for subsequent experiments.

Step B: A solution of adduct 3D2 (6.0 g, 17.06 mmol, 1 eq) in DMF (50mL) was cooled down to −45° C. Ethyl iodide (1.64 mL, 20.5 mmol, 1.2 eq)was then slowly added and the solution was stirred at −45° C. for 2 hand then at R.T. (16 h). Water was added and the mixture was extractedtwice with a mixture of ether/hexanes (1:1, 3×150 mL). The combinedorganic fractions were washed with water (2×), dried over MgSO₄,filtered and concentrated to afford approximately a 1:1 mixture of 3D3and 3D4 (S versus N alkylation) (6.1 g, 100%) as a yellow oil. Thismixture is used in the following step since only the S-alkylated analogcyclizes.

Step C: In a pre-heated sand bath (350° C.) a solution of compounds 3D3and 3D4 (6.1 g, 17.05 mmol, 1 eq.) in diphenyl ether (60 mL) was heateduntil the internal temperature reached 220° C., which was maintained for7 minutes. The solution was cooled to R.T. and the mixture loadeddirectly on a silica gel column, being eluted first with hexanes (1 L)to remove the diphenyl ether, and then 3% EtOAc/hexanes to afford thedesired quinoline 3D5 (2.76 g, 52%) as a pale yellow solid. The productwas suitable for use in the next step.

Step D: To a solution of quinoline 3D5 (2.76 g crude; 8.85 mmol; 1 eq)in THF (10 mL) and methanol (10 mL) at R.T. was added 1N NaOH (45 mL; 45mmol; 5.1 eq). The reaction was allowed to stir at reflux (85° C.) for24 h (monitored by HPLC). The mixture was acidified with 4N HCl andextracted with methylene chloride (3×). The organic fractions were driedover MgSO₄, filtered and concentrated to afford the quinoline acid 3D6(2.43 g, 97%) as a pale yellow solid. MS: (M+H)+; 284. This material wasused as is for the following reaction.

Step E: Compound 3D6 (2.43 g, 8.56 mmol) was put in diphenyl ether (20mL) and the heterogeneous mixture was heated to 250° C. for 12 minutesbefore being cooled. The mixture was directly transferred to a silicagel column and eluted first with hexanes (to remove diphenyl ether), andthen with 30% and 50% EtOAc/hexanes (R_(f)=0.48 in EtOAc/hexanes (1:1)).Evaporation of the solvent afforded the desired2-thioethyl-8-chloro-4-hydroxyquinoline 3D7 (1.25 g, 61%) as a paleyellow solid. MS: (M+H)+; 240, HPLC homogeneity=99%.

Example 3E Preparation of 2-(2-methylpropyl)-8-chloro-4-hydroxyquinoline(3E4)

Step A: Quinoline 3D7 (75 mg, 0.31 mmol, 1 eq) was dissolved inanhydrous DMF (6 mL) and then treated with K₂CO₃ (138 mg, 1 mmol, 3.2eq), To this suspension was added p-methoxybenzyl chloride (0.063 mL,0.47 mmol, 1.5 eq) and the mixture stirred at R.T. for 24 hours. Thereaction mixture was diluted with H₂O (20 mL) and extracted using amixture of diethyl ether/hexanes ((1:1), 3×40 mL). The organic phase wasdried (MgSO₄), filtered and concentrated to afford a pale yellow oilthat was purified on a silica gel column (eluent: 20% EtOAc/hexanes) togive the protected quinoline 3E1 (110 mg, 98%) as a clear oil.

Step B: To a solution of quinoline 3E1 (110 mg, 0.31 mmol, 1 eq) inmethanol (3 mL) and diether ether (1 mL) was added water (1 mL). Oxone(564 mg, 0.92 mmol, 3 eq) was then added and the mixture was stirred atR.T. for 2 h. The reaction mixture was diluted in water (20 mL) andextracted with methylene chloride (3×20 mL) and finally washed withbrine. The organic phase was dried (MgSO₄), filtered and concentrated toafford sulfone 3E2 (116 mg, 97%) as a pale yellow oil.

Step C: To a solution of sulfone 3E2 (116 mg, 0.296 mmol, 1 eq) in THF(3.0 mL) was added 2-methyl-1-propanol (2.0 mL, 10.83 mmol, 39 eq). Themixture was then treated with NaH (24 mg, 60%/oil, 0.592 mmol, 2 eq) andallowed to stir 45 min at R.T. The reaction was quenched by carefuladdition of H₂O (10 mL) and then extracted with diethyl ether (3×20 mL).The organic phases were combined, dried (MgSO₄), filtered andconcentrated. The product was purified over a silica gel column (eluent:5% EtOAc/hexanes) to afford the isobutoxy derivative 3E3 (87 mg, 79%) asa white solid.

Step D: To a solution of quinoline 3E3 (87 mg, 0.234 mmol) in methylenechloride (5 mL) was added slowly trifluoroacetic acid (5 mL) and themixture allowed to stir for 15 min at R.T. The mixture was concentratedand then purified over silica gel column (eluent: 10% EtOAc/hexanes toafford after drying hydroxyquinoline 3E4 (56 mg, 95%) as a white solid.MS: 252 (M+H)+.

Example 3F Preparation of 2-ethoxy-8-thiomethyl-4-hydroxyquinoline (3F4)

Step A: The imidate salt 3A2 (1.4 g, 7.2 mmol, 1 eq.) was combined with2-(methylthio)aniline 3F2 (0.96 g, 7.50 mmol, 1 eq.) in ethanol (15 mL)under an N₂ atmosphere. The reaction mixture was stirred at R.T. (1 h)and monitored by HPLC. The reaction mixture was concentrated and thenether was added and the mixture filtered. The solids were washed withether and the combined ether washes concentrated in vacuo. The resultingadduct 3F3 was obtained as a yellow oil (1.66 g, 82%) and used as is inthe next step. MS electrospray: (M+H)+; 282 and (M−H)−; 280.

Step B: The condensation product 3F3 (1.66 g, 5.90 mmol) was dissolvedin diphenyl ether (10 mL) and placed in a sand bath (300° C.). Theinternal temperature was monitored and maintained between 240-250° C.for 10 minutes. The mixture was cooled and then directly loaded on asilica gel column and eluted first with hexanes, then with 30%EtOAc/Hexanes and finally 50% EtOAc/hexanes. The product wasconcentrated and dried in vacuo to give the corresponding4-hydroxyquinoline derivative 3F4 as a yellow solid (0.735 g, 53%). MSelectrospray: (M+H)+; 236 and (M−H)−; 234.

Example 3G Preparation of2-ethoxy-8-(trimethylsilylethynyl)-4-hydroxyquinoline (3G3)

Step A: To 2-[(trimethylsilylethynyl)aniline] 3G1 (1.0 mL, 4.97 mmol, 1eq) was added imidate 3A2 (0.972 g, 4.97 mmol, 1 eq) in abs. ethanol (15mL) under a N₂ atmosphere. The mixture was stirred at R.T. for 48 h atwhich point the reaction was concentrated. The residue was taken up intodiethyl ether and the salts removed by filtration. The concentratedmixture was purified on a silica gel column (eluent: EtOAc/hexanes(5:95)) to afford adduct 3G2 (1.28 g, 78%) as an oil. MS: 332 (M+H)+.

Step B: Adduct 3G2 (928 mg, 2.79 mmol) was dissolved in diphenyl ether(10 mL) and placed in a pre-heated sand bath (300° C.). The internaltemperature was monitored and allowed to stay between 225° C.-235° C.for 7 minutes. The mixture was cooled and directly loaded on a silicagel column and eluted with hexanes to remove diphenyl ether, followed bya gradient of 30% to 50% EtOAc/hexanes. Concentration and drying invacuo afforded the desired quinoline 3G3 (463 mg, 58%) as a beige solid.MS: 286 (M+H)+.

Note: Subsequent brosylate displacement with this TMS protected ethynylquinoline 3G3 on the macrocyclic tripeptide gives directly the desiredhydroxyquinoline analog with loss of the TMS group in situ.

Example 3H Preparation of 2-ethoxy-8-methoxy-4-hydroxyquinoline (3H3)

Step A and B: Beginning with ortho-anisidine 3H1 and following the sameprotocol as outlined in previous examples 3F and 3G, the desired8-methoxyquinoline derivative 3H3 was obtained in 38% overall yield as apale yellow solid. MS: 220 (M+H)+.

Example 3I Preparation of 2-ethoxy-8-bromo-7-methoxy-4-hydroxyquinoline(3I2)

Step A: To 2-bromo-3-aminoanisole 2B4 (750 mg, 3.7 mmol, 1 eq) was addedimidate 3A2 (0.73 g, 3.7 mmol, 1 eq) in ethanol (7 mL) under a N₂atmosphere. The mixture was stirred at R.T. for 24 h at which point thereaction was concentrated and purified directly on a silica gel column(eluent: EtOAc/Hexanes (1:9)) to afford adduct 311 (1.12 g, 88%) as apale yellow oil. MS: 344 (M+H)+ and 346 (MH+2)+.

Step B: Adduct 311 (1.12 g, 3.25 mmol) was dissolved in diphenyl ether(10 mL) and placed in a pre-heated sand bath (300° C.). The internaltemperature was monitored and allowed to stay between 240° C.-250° C.for 8 minutes. The mixture was directly loaded on a silica gel columnand eluted with hexanes to remove diphenyl ether, followed by a gradientof 30% to 50% EtOAc/hexanes: (Rf=0.25 in 1:1 EtOAc/hexanes).Concentration and drying in vacuo afforded the desired quinoline 312(734 mg, 76%) as a white solid. MS: 298 (M+H)+ and 300 (MH+2)+.

Example 3J Preparation of 8-bromo-7-methoxy-4-hydroxyquinolone (3J1)

Step A: To 2-bromo-3-aminoanisole 2B4 (378 mg, 1.87 mmol, 1 eq) andmalonic acid (194 mg, 1.87 mmol) was added phosphorous oxychloride (175μL, 1.87 mmol). The reaction was placed in a pre-heated bath (95° C.)and stirred for 30 minutes. The mixture was cooled, and diluted with icewater and stirred ca. 2 h to afford a grey solid. This was filtered andwashed with water until free of acid and then taken up into 1N NaOH.Insoluble material was removed by filtration and the resulting aqueoussolution was treated with EtOH (8 mL) before adjusting the pH to 5-6with 1N HCl (aq). The desired product was filtered and dried to affordquinolone 3J1 (0.12 g, 24%) as a yellow solid.

MS: 270.0 (M+H)+ and 291.9 (M+Na)+.

Example 3K Preparation of 5-ethoxythieno[3.2-b]pyridin-7-ol (3K3)

Step A: To available thiophen-3-ylamine 3K1 (0.50 g, 5.04 mmol) wasadded imidate 3A2 (1.08 g, 5.5 mmol) in ethanol (10 mL) under a N₂atmosphere. The mixture was stirred at R.T. for 3 h at which point thereaction was concentrated. To the residue was added ether, and thesuspension filtered and washed with ether to afford adduct 3K2(1.0 g,82%). This material was sufficiently clean to be used in the subsequentstep.

MS: 242.1 (MH)+.

Step B: Adduct 3K₂ (1.0 g, 4.14 mmol) was dissolved in diphenyl ether (5mL) and placed in a pre-heated sand bath (300° C.). The internaltemperature was monitored and allowed to stay between 210° C.-225° C.for 7 minutes. The mixture was directly loaded on a silica gel columnand eluted with hexanes to remove diphenyl ether, followed by a gradientof 30% EtOAc/hexane to neat EtOAc. Concentration and drying in vacuoafforded the desired thieno[3.2-b]pyridinol 3K₃ (200 mg, 25%) as a brownsolid. MS: 196 (MH+)+.

Example 3L Preparation of 5-ethoxy-3-methylthieno[3.2-b]pyridin-7-ol(3L4)

The method of J. M. Barker, P. R. Huddleston, M. L. Wood, Synth. Commun.(1995) 25(23): 3729 was followed for steps A and B.

Step A: 1M NaOH (20 mL) was added to compound 3L1 (1.5 g, 8.76 mmol).The mixture was heated at reflux for 3 h, then cooled to RT, acidifiedto pH=1 with conc. HCl. The solution was filtered and rinsed once withminimal amount of hexanes. The solid that was obtained was dissolved inacetone/MeOH and dried over sodium sulfate, filtered and concentrated toobtain compound 3L2 as an off-white solid (1.37 g, 99%) which wasimmediately employed in subsequent step. LC-MS t_(R)=2.88 min,ES-=155.9.

Step B: To a suspension of amino acid 3L2 (1.38 g, 8.78 mmol) in i-PrOH(20 mL) was added oxalic acid (1.00 g, 11.11 mmol). This opaque solutionwas gently warmed to about 40° C. for 1 h until no starting material wasobserved by LC-MS. The mixture was cooled to RT, the solvent removed andthe crude residue purified by flash column:chromatography (4:1 to 1:1 to1:4 hex:ETOAc) to yield the desired amine 3L3 (350 mg, 27%). ¹H NMR (400MHz, CDCl₃); 7.20 (br s, 2H), 6.86 (s, 1H), 6.00 (s, 1H), 1.99 (s, 3H).

Step C: Amine 3L3 was converted to compound 3L4 using a procedureanalogous to that described in Example 3K.

Example 4 Synthesis of Compound 104 from Table 1

Step A: To a solution of the macrocyclic brosylate intermediateINRF12Brs (50 mg, 0.070 mmol, 1.0 eq.), dissolved in NMP (4 mL) wasadded the hydroxyquinoline 3A5 (15.7 mg, 0.070 mmol, 1.0 eq.) and cesiumcarbonate (25.09 mg, 0.077 mmol, 1.1 eq.). The mixture was heated at 70°C. for 16 hours. After the complete conversion of starting material toproducts, the reaction mixture was diluted with EtOAc and washed withH₂O (2×), saturated aq. NaHCO₃ (2×), and brine (1×). The organic layerwas dried over anhydrous MgSO₄, filtered and evaporated to dryness.Product 41 (48.8 mg, 100%) was sufficiently pure to be used directly inthe following step.

Step B: The methyl ester 41 (47.8 mg, 1.0 mmol) was dissolved in asolution of THF/MeOH/H₂O (2:1:1, 1.2 mL) and saponified with 1N NaOH(0.56 mL, 0.56 mmol, 8 eq.). The hydrolysis reaction was carried outover 5 h at RT. Thereafter, the solution was evaporated to dryness togive an off-white solid. This material was dissolved in acetic acid andpurified by preparative HPLC (AcCN/H₂O/TFA). Pure fractions werecombined, frozen, and lyophilized to afford compound 104 as a whitesolid (12.3 mg; 26% yield), 97% homogeneity by analytical HPLC.

¹H NMR (400 MHz, DMSO-d₆): δ 8.51 (s, 1H), 7.93 (d, J=8 Hz, 1H), 7.73(d, J=8 Hz, 1H), 7.21-7.15 (m, 2H), 6.56 (s, 1H), 5.50-5.35 (m, 2H),5.20 (bdd, J=9.4, 9.4 Hz, 1H), 4.50-4.40 (m, 4H), 4.40-4.30 (m, 1H),4.1-3.8 (m, 2H underwater), 2.30-2.18 (m, 1H), 2.15-2.05 (m, 1H),1.75-1.60 (m, 2H), 1.60-1.35 (m, 11H), 1.34 (t, J=7.1 Hz, 3H), 1.32-1.0(m, 8H).

Example 5 Synthesis of Compounds 118 and 124 from Table 1

Step A: To a solution of the macrocyclic brosylate intermediateINRF12Brs (150 mg, 0.212 mmol, 1.0 eq.), dissolved in NMP (5 mL) wasadded the hydroxyquinoline 3F3 (50 mg, 0.212 mmol, 1.0 eq.) and cesiumcarbonate (82.8 mg, 0.254 mmol, 1.2 eq.). The mixture was heated at 70°C. for 16 hours. After the complete conversion of starting material toproducts, the reaction mixture was diluted with EtOAc and washed withH₂O (2×), saturated aq. NaHCO₃ (2×), and brine (1×). The organic layerwas dried over anhydrous MgSO₄, filtered and evaporated to dryness.Product 51 (88 mg, 59%) was sufficiently clean to be used directly inthe following step. MS: (MH)+; 709.3, (M−H)−; 707.3.

Step B: The methyl ester 51 (88 mg, 0.124 mmol) was dissolved in asolution of THF/MeOH (2 mL each) and then treated with 1N NaOH (2.0 mL,2.0 mmol). The hydrolysis reaction was carried out over 18 h at RT.Thereafter, the solution was evaporated to dryness to give an off-whitesolid. This material was dissolved in acetic acid and purified bypreparative HPLC (AcCN/H₂O/TFA). Pure fractions were combined, frozen,and lyophilized to afford compound 118 as a beige solid (57 mg; 66%yield). ¹H NMR (400 MHz, DMSO-d₆): δ 8.57 (s, 1H), 7.76 (d, 1H, J=8 Hz),7.35 (d, 1H, J=8 Hz), 7.27-7.20 (m, 2H), 6.55 (s, 1H), 5.54-5.47 (m,1H), 5.41 (brs, 1H), 5.26 (t, 1H, J=9 Hz), 4.60-4.38 (m, 4H), 4.09 (brs,1H), 3.84 (brd, 1H, J=9 Hz), 3.16 (s, 1H), 2.55 (m, 1H), 2.43 (s, 3H),2.31-2.14 (m, 2H), 1.80-1.05 (m, 23H); MS (MH)⁺; 695.4, (M−H)⁻; 693.3,analytical HPLC homogeneity=98.7%.

Step C: To a solution of compound 118 (35 mg, 0.050 mmol) in a 1:1mixture of MeOH/water (1.5 mL each) was added Oxone® (0.154 g, 0.25mmol) all at once. The reaction was stirred at RT for 7 h before beingconcentrated. The residue was taken up into methylene chloride (20 mL)and washed with water (2×20 mL) and sat. brine (3×20 mL). The organicphase was dried over MgSO₄, filtered and concentrated to give thedesired crude product. The material was dissolved in AcOH and purifiedby preparative HPLC (Combiprep ODS-AQ, 20×50 mm) to give the desiredcompound 124 (2.5 mg, 7%) as a white solid. MS (MH)+; 727.3, (M−H)−;725.3, analytical HPLC homogeneity=100%.

¹H NMR (400 MHz, DMSO-d₆): δ 8.59 (s, 1H), 8.36 (d, J=8 Hz, 1H), 8.25(d, J=7 Hz, 1H), 7.49 (dd, J=8 Hz, 1H), 7.25 (d, J=7 Hz, 1H), 6.73 (s,1H), 5.55-5.47 (m, 2H), 5.25 (dd, J=9 Hz, 1H), 4.58-4.47 (m, 3H),4.46-4.39 (m, 1H), 4.10-4.01 (m, 2H), 3.83 (d, J=9 Hz, 1H), 3.53 (s,3H), 2.35-2.26 (m, 1H), 2.19-2.10 (m, 1H), 1.77-1.66 (m, 2H), 1.56-1.38(m, 13H), 1.38-1.25 (m, 7H), 1.24-1.08 (m, 2H).

Example 6 Synthesis of Cyclopropylsulfonamide Intermediate 64

Step A: A dry 3 L 3-neck flask equipped with a magnetic stir bar,addition funnel and argon inlet was flushed with argon, then chargedwith 3-chloropropanesulfonyl chloride 61 (100.48 g, 0.57 mol, 1.0 eq).Anhydrous dichloromethane (900 mL) was transferred into the flask viacannula, the mixture was cooled in an ice/water bath and tert-butylamine(72 mL, 0.68 mol, 1.2 eq) was added. The mixture was stirred 15 minutesthen a solution of triethylamine (158 mL, 1.13 mol, 2.0 eq) in anhydrousdichloromethane (100 mL) was added dropwise over 45 minutes and stirringwas continued for 1 h. The mixture was diluted with dichloromethane (500mL) and washed with 1N HCl (3×400 ml) and brine. The organic layer wasdried over sodium sulfate, filtered and evaporated to dryness to givecompound 62 as an orange-beige solid (107.04 g, 88% yield). ¹H NMR(CDCl₃, 400 MHz): 64.46 (s, 1H), 3.71 (tr, 2H), 3.25 (tr, 2H), 2.31 (m,2H), 1.41 (s, 9H).

Step B: A dry 5 L 3-neck flask equipped with a magnetic stir bar, argoninlet and 2 addition funnels was flushed with argon and anhydrous THF(1.5 L) was transferred into the flask via cannula and cooled to −78° C.Compound 62 (96.73 g, 0.453 mol, 1.0 eq) was dissolved in anhydrous THF(390 mL) and the solution was transferred into one of the additionfunnels. n-Butyllithium solution (2.5 M in hexanes, 390 mL, 0.975 mol,2.15 eq) was transferred to the other addition funnel and the solutionsin the addition funnels were added to the flask simultaneously over 4hours. When addition was complete, the mixture was allowed to warm toroom temperature. Once the internal temperature reached ˜0° C., thereaction was quenched by dropwise addition of saturated NH₄Cl solution(200 mL). The THF was removed under vacuum and the residue was dilutedwith CH₂Cl₂ (2 L) and water (1 L). The layers were separated and theorganic layer was washed with water (2×1 L) and brine (800 mL), driedover sodium sulfate, filtered and evaporated to dryness. Compound 63 wasobtained as an orange-beige solid (77.32 g, 96% yield). ¹H NMR (CDCl₃,400 MHz): 64.25 (s, 1H), 2.48 (m, 1H), 1.42 (s, 9H), 1.19 (m), 1.01 (m).

Step C: A 2 L flask equipped with a magnetic stir bar and condenser wascharged with Compound 63 (82.53 g, 0.466 mol, 1.0 eq), dichloromethane(400 mL) and trifluoroacetic acid (460 mL, 5.97 mol, 13 eq). The mixturewas heated to reflux for 2 h, allowed to cool, and evaporated andco-evaporated several times with CH₂Cl₂ to remove most of the TFA. Thecrude product was dissolved in 95:5 CH₂Cl₂:MeOH and NH₄OH and waspurified by silica gel column chromatography (94:5:1 CH₂Cl₂:MeOH:NH₄OH).Compound 64 was obtained as a beige solid (46.38 g, 78% yield). ¹H NMR(DMSO-d₆, 400 MHz): δ 6.79 (s, 2H), 2.54 (1H, under DMSO peak), 0.92(4H).

Example 7 Synthesis of Compound 136 from Table 1

Compound 101 (Table 1) (24 mg, 0.032 mmol), prepared from INRF12Brs and2-ethoxy-8-bromo-7-methoxy-4-hydroxy quinoline (312; Example 31) using aprocedure analogous to the one described in Example 4, was combined withHATU (14 mg, 0.038 mmol) in anhydrous DMF (4 mL). The solution wasstirred at R.T. before DIPEA (22 μL, 0.13 mmol) was added dropwise overca. 1 min. The mixture was stirred for 1 h at R.T. and analyzed byanalytical HPLC for the formation of the activated ester. A solution ofmethanesulfonamide (12 mg, 0.13 mmol), DMAP (15 mg, 0.12 mmol) and DBU(19 μL, 0.13 mmol) were added in DMF (1 mL). The reaction mixture wasstirred 48 h at R.T. before being concentrated. The reaction mixture waspoured into EtOAc (50 mL) and washed with sat. NaHCO₃ (aq) and sat.brine, before being dried over Na₂SO₄, filtered and concentrated. Theresidue was reconstituted in DMSO and purified by preparative HPLC.Lyophilization of pure fractions gave the Compound 136 (7.8 mg, 29%) asa white amorphous solid. MS: 834.2 (M+H)+ and 836.2 (MH+2)+.

¹H NMR (400 MHz, DMSO-d₆) δ 11.17 (s, 1H), 8.95 (s, 1H), 7.99 (d, J=9Hz, 1H), 7.33 (d, J=7 Hz, 1H), 7.15 (d, J=9 Hz, 1H), 6.49 (s, 1H), 5.57(dd, J=9 Hz, 1H), 5.46 (bs, 1H), 5.14 (dd, J=9.5, 9.5 Hz, 1H), 4.63-4.55(m, 1H), 4.50 (q, J=7 Hz, 2H), 4.44-4.33 (m, 2H), 4.09-3.99 (m, 2H),3.94 (s, 3H), 3.83 (d, J=8.6 Hz, 1H), 3.16 (s, 3H), 2.65-2.53 (m, 1H),2.35-2.25 (m, 1H), 1.77-1.45 (m, 14H), 1.39 (t, J=7 Hz, 3H), 1.38-1.08(m, 7H). Compounds of formula (I) wherein R¹ is NHSO₂R¹¹ and R¹¹ iscyclopropyl may be prepared by the method of Example 7 but usingcyclopropylsulfonamide 64 in place of methanesulfonamide.

Example 8 Synthesis of Compound 135 Table 1 Preparation of SulfamideIntermediate 83

Step A: Reagent 81 (0.3 g, 0.99 mmol) [prepared according to Winum, J-Y;Toupet, L; Barragan, V; Dewynter, G; Montero, J-L., Org. Lett., 14(3),2241-2243 (2001)] was suspended in CH₂Cl₂, morpholine (0.086 mL, 0.99mmol) was added and the mixture was stirred for 5 h. The reaction wasfollowed by TLC. On completion the reaction mixture was directlyadsorbed on silica gel and the product was eluted with 6% MeOH in CHCl₃to afford 0.258 g (98%) of compound 82 as a white solid.

Step B: Compound 82 (0.150 g, 0.56 mmol) was dissolved in CH₂Cl₂ (5 mL)and treated with TFA (1 mL). The reaction was stirred for 4 h andmonitored by TLC. Upon completion, the solvent was evaporated and theresidue directly adsorbed on the silica gel and eluted with 5% MeOH inCHCl₃ to afford 0.075 g (80.2%) of compound 83 as a white solid.

Synthesis of Compound 135 (Table 1)

Compound 101 (24 mg, 0.032 mmol) (described in Example 7) was combinedwith HATU (14 mg, 0.038 mmol) in anhydrous DMF (4 mL). The solution wasstirred at R.T. before DIPEA (22 μL, 0.13 mmol) was added dropwise overca. 1 min. The mixture was stirred for 1 h at R.T. and analyzed byanalytical HPLC for the formation of the activated ester. A solution ofcompound 83 (21 mg, 0.13 mmol), DMAP (15 mg, 0.12 mmol) and DBU (19 μL,0.13 mmol) were added in DMF (1 mL). The reaction mixture was stirred 48h at R.T. before being concentrated. The reaction mixture was pouredinto EtOAc (50 mL) and washed with sat. NaHCO₃ (aq) and sat. brine,before being dried over Na₂SO₄, filtered and concentrated. The residuewas reconstituted in DMSO and purified by preparative HPLC.Lyophilization of pure fractions gave the sulfamide derivative Compound135 (7.5 mg, 26%) as a white amorphous solid. MS: 905.3 (M+H)⁺ and 907.3(MH+2)⁺.

¹H NMR (400 MHz, DMSO-d₆) δ10.93 (s, 1H), 8.85 (s, 1H), 7.99 (d, J=9 Hz,1H), 7.33 (d, J=6.5 Hz, 1H), 7.15 (d, J=9 Hz, 1H), 6.48 (s, 1H),5.68-5.56 (m, 1H), 5.50-5.42 (m, 1H), 5.13-5.10 (m, 1H), 4.59 (bd, J=8Hz, 1H), 4.55-4.43 (m, 3H), 4.40 (dd, J=7.0, 7.0 Hz, 1H), 4.10-4.0 (m,1H), 3.94 (s, 3H), 3.84 (bd, J=9 Hz, 1H), 3.60-3.53 (m, 4H), 3.17-3.06(m, 4H), 2.70-2.57 (m, 1H), 2.35-2.23 (m, 1H), 1.78-1.48 (m, 14H), 1.39(t, 3H), 1.34-1.10 (m, 7H).

Example 9 Parallel Synthesis of Compounds Exemplified in Table 2

Step A: A series of 8-mL vials were disposed in a reaction block. Ineach vial was successively added the brosylate (0.07 mmol, 48.9 mg), NMP(1 mL), the building block R²—OH (0.08 mmol) and cesium carbonate (0.12mmol, 37.6 mg). The closed vials were placed in an oil bath at 70° C.for 18 h. Each reaction mixture was diluted with EtOAc (10 mL) andsuccessively washed with water, saturated aqueous NaHCO₃ and brine.After the usual treatment (MgSO₄, filtration and concentration), thecrude materials were purified by flash chromatography (SiO₂, elutionwith hexane-EtOAc, 1:1 to 4:6 for diverse 4-hydroxy quinolines). Thepurified compounds were transferred into 8-mL vials, vacuum centrifugedto remove the solvent and weighed (the amount of desired material variedbetween 0.02 and 0.025 mmol).

Step B: Removal of the Boc protecting group. All the vials were treatedwith 4M HCl in dioxane (1 mL) for 1 h and vacuum centrifuged to removethe volatiles.

Step C: In vials was added the corresponding R³-carbonate (approximately2 eq. based on the yield of the first step) (0.04 mmol) and DIPEA (0.09mmol, 16 μL) in DCE (500 μL) or the corresponding R³-chloroformate(approximately 3 eq. based on the yield of the first step) (0.06 mmol)and DIPEA (0.09 mmol, 16 μL) in DCE (500 μL). The reactions were allowedto proceed overnight. All vials were then vacuum centrifuged to removethe volatiles.

Step D: All reactions were diluted with 1 mL of THF and 500 μL of MeOH.A solution of 500 μL of 2N aq. LiOH (1 mmol) was then added and thesolutions were allowed to react overnight. The reaction mixtures werethen vacuum centrifuged to remove the volatile material, diluted with500 μL of DMSO and neutralized by the addition of 400 μL of AcOH.

Step E: All compounds were purified by semi-prep reversed-phase HPLC(Symmetry column 5 cm×19 cm, CH₃CN/H₂O 0.06% TFA gradient).

Example 10 Compound 116 Table 1

¹H NMR (400 MHz, DMSO-d₆) δ12.25 (bs, 1H), 8.57 (s, 1H), 8.12 (d, J=8Hz, 1H), 7.88 (d, J=7 Hz, 1H), 7.38 (dd, J=8, 8 Hz, 1H), 7.24 (d, J=7Hz, 1H), 7.14 (s, 1H), 5.62-5.45 (m, 2H), 5.26 (dd, J=9, 9 Hz, 1H),4.70-4.53 (m, 2H), 4.41 (dd, J=8 Hz, 1H), 4.13-4.04 (m, 2H), 3.86 (m,2H), 2.95 (q, J=8 Hz, 2H), 2.62-2.50 (m, 1H), 2.37-2.25 (m, 1H),2.22-2.12 (m, 1H), 1.80-1.60 (m, 3H), 1.60-1.36 (m, 13H), 1.34 (t, 3H),1.25-1.08 (m, 2H).

Compound 205 Table 2

¹H NMR (400 MHz, DMSO-d₆) δ 12.2 (bs, 1H), 8.58 (S, 1H), 7.81 (d, J=8Hz, 1H), 7.50-7.42 (m, 2H), 7.31-7.23 (m, 1H), 6.92 (s, 1H), 5.55-5.50(m, 1H), 5.48-5.43 (m, 1H), 5.26 (dd, J=8, 8 Hz, 1H), 4.71-4.65 (m, 1H),4.45 (q, J=7 Hz, 2H), 4.45-4.35 (m, 1H), 4.13-4.05 (m, 1H), 3.85 (dd,J=8, 8 Hz, 1H), 3.65-3.52 (m, 4H), 3.48 (d, J=10 Hz, 1H), 2.62-2.53 (m,1H), 2.48-2.42 (m, 1H), 2.35-2.25 (m, 1H), 2.16 (dd, J=9 Hz, 1H),1.93-1.80 (m, 1H), 1.79-1.65 (m, 3H), 1.60-1.51 (m, 1H), 1.50-1.42 (m,3H), 1.37 (t, J=7 Hz, 3H), 1.36-1.05 (m, 5H).

Compound 306 Table 3

¹H NMR (400 MHz, DMSO-d₆) δ12.4 (bs, 1H), 8.46 (s, 1H), 7.84 (d, J=8 Hz,1H), 7.49 (d, J=7 Hz, 1H), 7.18 (dd, J=7.5, 7.5 Hz, 1H), 7.075 (d, J=7Hz, 1H), 6.50 (s, 1H), 5.37 (bs, 1H), 4.71-4.6 (m, 2H), 4.47 (q, J=7 Hz,2H), 4.44-4.35 (m, 2H), 4.26-4.15 (m, 1H), 3.82 (bd, J=8 Hz, 1H), 2.57(s, 3H), 2.45-2.35 (m, 1H), 2.35-2.23 (m, 1H), 1.80-1.00 (m, 25H), 1.39(t, J=7 Hz, 3H).

Example 11 Synthesis of Compound 402 Table 4

Step A. To a solution of the macrocyclic brosylate intermediate INRF12Br(318 mg, 0.448 mmol, 1.0 eq.), dissolved in NMP (10 mL) was added thehydroxyquinolone 3J1 (121 mg, 0.448 mmol, 1.0 eq.) and cesium carbonate(153 mg, 0.47 mmol, 1.1 eq.). The mixture was heated at 70° C. for 24hours. After the complete conversion of starting material to products,the reaction mixture was diluted with EtOAc and washed with H₂O (2×),saturated aq. NaHCO₃ (2×), and brine (1×). The organic layer was driedover anhydrous Na₂SO₄, filtered and evaporated to dryness. The crudereaction material was purified by chromatography (SiO₂, EtOAc) to giveproduct 11.1 (143 mg, 43%) as a white solid. MS: 743.3 (M+H)+ and 745.3(MH+2)+.

Step B. Alkylation of the quinolone adduct 11.1 (70 mg, 0.094 mmol) wasaccomplished in acetonitrile (5 mL) with MeI (58 μL, 0.93 mmol) withpotassium carbonate (26 mg). The reaction was stirred at 70° C. for 5 hbefore a second amount of MeI was added (58 μL, 0.93 mmol). The mixturewas stirred a further 16h before being concentrated. The residue wassuspended in chloroform, filtered to remove salts, and concentrated togive a mixture of compounds 11.2 and 11.3 as a white solid. MS: 757.3(M+H)+, (MH+2)+, 759.3. This mixture was used as such in the next step.

Step C. The methyl esters 11.2 and 11.3 (71 mg, 0.094 mmol) weredissolved in a solution of THF/MeOH/H₂O (2:1:1, 2 mL) and saponifiedwith 1N NaOH (0.75 mL, 0.75 mmol, 8 eq.). The hydrolysis reaction wascarried out over 5h at RT. Thereafter, the solution was evaporated todryness to give an off-white solid. This material was dissolved inacetic acid and purified by preparative HPLC (AcCN/H₂O/TFA). Purefractions were combined, frozen, and lyophilized to afford the cyclictripeptide Cpd 402 as a white solid (4.3 mg; 22% yield), 98% homogeneityby analytical HPLC. MS: 743.2 (M+H)+ and 745.2 (MH+2)+.

¹H NMR (400 MHz, DMSO-d₆) δ12.21 (bs, 1H), 8.57 (s, 1H), 7.88 (d, J=9Hz, 1H), 7.21 (d, J=7 Hz, 1H), 6.98 (d, J=9 Hz, 1H), 6.03 (s, 1H), 5.50(dd, J=8, 8 Hz, 1H), 5.34 (bs, 1H), 5.26 (dd, J=10 Hz, 1H), 4.53-4.36(m, 3H), 4.09-4.00 (m, 1H), 3.92 (s, 3H), 3.80-3.73 (m, 1H), 3.70 (s,3H), 2.47-2.39 (m, 1H), 2.33-2.20 (m, 1H), 2.19-2.09 (dd, J=9, 9 Hz,1H), 1.79-1.56 (m, 3H), 1.56-1.23 (m, 15H), 1.23-1.05 (m, 2H).

Example 12 NS3-NS4A Protease Assay

The enzymatic assay used to evaluate the present compound is describedin WO 00/09543 and WO 00/59929.

Example 13 Cell-Based Luciferase Reporter HCV RNA Replication Assay

The cell-based HCV RNA replication assay used to evaluate the presentcompounds is described as follows:

Cell Culture:

Huh-7 cells that stably maintained a subgenomic HCV replicon containingthe luciferase-FMV2A-neomycin phosphotransferase fusion gene wereestablished as previously described (Lohman et al., 1999. Science 285:110-113) and designated as MP-1 cells. MP-1 cells are maintained inDulbecco's Modified Earle Medium (DMEM) supplemented with 10% FBS and0.25 mg/mL neomycin (standard medium). The cells are passaged bytrypsinization and frozen in 90% FBS/10% DMSO. During the assay, DMEMmedium supplemented with 10% FBS, containing 0.5% DMSO and lackingneomycin used (Assay medium). The day of the assay, MP-1 cells aretrypsinized and diluted to 100 000 cells/mL in assay medium. 100 μL isdistributed into each well of a black 96-well ViewPlate™ (Packard). Theplate is then incubated at 37° C. with 5% CO₂ for two hours.

Reagents and Materials

Product Company Catalog # Storage DMEM Wisent Inc. 10013CV  4° C. DMSOSigma D-2650 RT Dulbecco's PBS Gibco-BRL 14190-136 RT Fetal Bovine SerumBio-Whittaker 14-901F −20° C./4° C. Geneticin (G418) Gibco-BRL 10131-027−20° C./4° C. Trypsin-EDTA Gibco-BRL 25300-054 −20° C./4° C.ViewPlate ™-96, Black Packard 6005182 RT Backing tape, Black Packard6005189 RT PVDF 0.22 μm Filter Millipore SLGV025LS RT Unit Deep-WellTiter Plate Beckman 267007 RT Polypropylene

Preparation of Test Compound:

The test compound in 100% DMSO was first diluted in assay medium to afinal DMSO concentration of 0.5%. The solution was sonicated for 15 minand filtered through a 0.22 μM Millipore Filter unit. Into column 3 of aPolypropylene Deep-Well Titer Plate, the appropriate volume istransferred into assay medium to obtain the starting concentration (2×)to be tested. In columns 2 and, 4 to 12 add 200 μL of assay medium(containing 0.5% DMSO). Serial dilutions (1/2) are prepared bytransferring 200 μL from column 3 to column 4, then from column 4 tocolumn 5, serially through to column 11. Columns 2 and 12 are the noinhibition controls.

Addition of Test Compound to Cells:

A volume of 100 μL from each well of the compound dilution plate istransferred to a corresponding well of the Cell Plate (Two columns willbe used as the “No inhibition control”; ten [10] columns are used forthe dose response). The cell culture plate was incubated at 37° C. with5% CO₂ for 72 hours.

Luciferase Assay:

Following the 72h incubation period, the medium is aspirated from the96-well assay plate and a volume of 100 μL of 1×Glo Lysis Buffer(Promega) previously warmed to room temperature was added to each well.The plate was incubated at room temperature for 10 min with occasionalshaking. A black tape was put at the bottom of the plate. 100 μL ofBright-Glo luciferase substrate (Promega) previously warmed to roomtemperature was added to each well followed by gentle mixing. Theluminescence was determined on a Packard TopCount instrument using theData Mode Luminescence (CPS) with a count delay of 1 min and a counttime of 2 sec.

Product Company Catalog # Storage Glo Lysis Buffer Promega E266A  4° C.Bright-Glo Luciferase Assay System Promega E2620 −20° C.

The luminescence determination (CPS) in each well of the culture platewas a measure of the amount of HCV RNA replication in the presence ofvarious concentrations of inhibitor. The % inhibition was calculatedwith the following equation:

%inhibition=100−[CPS(inhibitor)/CPS(control)×100]

A non-linear curve fit with the Hill model was applied to theinhibition-concentration data, and the 50% effective concentration(EC₅₀) was calculated by the use of SAS software (Statistical Software;SAS Institute, Inc. Cary, N.C.).

When the compounds of this invention are evaluated in the precedingenzymatic and cell based assays, the compounds are found to be highlyactive. More specifically, the compounds have IC₅₀ values below 100 nMin the NS3-NS4A protease assay, and EC₅₀ values below 100 nM in thecell-based luciferase reporter HCV RNA replication assay.

Example 14 Specificity Assays

The specificity assays used to evaluate the selectivity of this compoundare described in WO 00/09543.

When the compounds are evaluated in the specificity assays, thecompounds of formula 1 are found to be selective in that they do notshow significant inhibition (no measurable activity at concentrations upto 30 μM) in the Human Leukocyte Elastase and Cathepsin B assays.

Tables of Compounds

The following tables list compounds representative of the invention. Allcompounds listed in Tables 1 to 5 were found to have IC₅₀ values below100 nM in the NS3-NS4A protease assay of Example 12. In addition, manyof the compounds listed in Table 1 have EC₅₀ values below 100 nM in thecell-based luciferase reporter HCV RNA replication assay of Example 13.Retention times (t_(R)) for each compound were measured using thestandard analytical HPLC conditions described in the Examples. As iswell known to one skilled in the art, retention time values aresensitive to the specific measurement conditions. Therefore, even ifidentical conditions of solvent, flow rate, linear gradient, and thelike are used, the retention time values may vary when measured, forexample, on different HPLC instruments. Even when measured on the sameinstrument, the values may vary when measured, for example, usingdifferent individual HPLC columns, or, when measured on the sameinstrument and the same individual column, the values may vary, forexample, between individual measurements taken on different occasions.

TABLE 1

Cpd R²² R²¹ R²⁰ R¹ t_(R) (MH)⁺ (MH + 2)⁺ 101 OMe Br OEt OH 6.9 757.3759.3 102 OMe Me OEt OH 5.6 693.4 103 H Br OEt OH 7.4 725.2 727.2 (M −H)⁻ (M − H + 2)⁻ 104 H Cl OEt OH 7.2 685.3 685.3 105 H Br OMe OH 6.0713.2 715.2 106 H Cl OMe OH 5.8 669.3 671.3 107 H F OMe OH 5.2 653.3 108OMe Me OMe OH 4.6 679.3 109 OMe Me H OH 4.3 649.3 110 H Cl H OH 4.7639.2 641.2 111 H Br H OH 5.1 683.2 685.2 112 H Cl Me OH 5.1 653.3 655.3113 H F OEt OH 7.4 667.3 114 H Me OEt OH 6.8 663.4 115 H Br Me OH 5.2697.3 699.3 116 H Cl Et OH 5.4 667.3 669.3 117 H Cl —CH(CH₃)₂ OH 5.7681.3 683.3 118 H SMe OEt OH 8.0 695.4 119 H Me O—CH(CH₃)₂ OH 7.0 677.3120 OMe Me O—CH(CH₃)₂ OH 6.5 707.4 121 H Cl O—CH(CH₃)₂ OH 8.4 697.4699.4 122 H Br O—CH(CH₃)₂ OH 8.6 741.3 743.3 123 H F O—CH(CH₃)₂ OH 7.7681.4 124 H SO₂Me OEt OH 6.8 727.3 125 H Cl SMe OH 8.0 685.3 687.3 126OMe Br OMe OH 7.5 743.3 745.3 127 H SMe H OH 5.7 651.3 128 H SOMe H OH5.3 667.2 129 H SO₂Me H OH 5.6 683.2 130 H Cl SEt OH 8.1 699.3 701.3 131H Cl OCH₂CH(CH₃)₂ OH 8.5 711.3 713.3 132 H Cl OCH₂CH₂CH₃ OH 8.1 697.3699.3 133 H OMe OEt OH 5.3 679.4 134 H —C≡CH OEt OH 7.1 673.4 135 OMe BrOEt

8.1 905.3 907.3 136 OMe Br OEt

7.9 834.2 836.2 137 H Cl SCH₂CH₂CH₃ OH 8.3 713.3 715.3 138 H ClSCH(CH₃)₂ OH 8.3 713.3 715.3 139 H Cl OCH₂C(CH₃)₃ OH 8.5 725.3 727.3 140H Cl OCH₂CH₂CF₃ OH 8.0 751.3 753.3 141 H Cl

OH 8.4 723.3 721.3 142 H Cl

OH 7.6 693.2 695.2 143 H Cl

OH 7.9 695.2 697.2 144 H Cl

OH 5.7 726.3 728.3 145 H Cl —C≡CH OH 6.0 663.2 665.2 146 OMe Br—OCH₂C≡CH OH 7.4 767.2 769.2 147 OMe Br —OCH₂CH≡CH₂ OH 7.6 769.2 771.2148 H Cl

OH 5.9 760.3 149 OMe CH₃ —OCH₂CH≡CH₂ OH 6.6 705.3 707.3 150 OMe CH₃—OCH₂C≡CH OH 7.2 703.3 705.3 151 OMe Cl —OEt OH 6.9 713.3 715.3 152 OMeCl —O—CH(CH₃)₂ OH 7.2 727.3 153 OMe Cl —OMe OH 6.7 699.3 701.3 154 OMeCl —OCH₂CH≡CH₂ OH 7.4 725.3 155 OMe Cl —OCH₂C≡CH OH 7.3 723.3 725.3 156OMe Br —OCH₂C≡CCH₃ OH 7.6 781.2 783.2 157 H Cl

OH 5.8 749.2 158 H Cl

OH 7.3 713.3 715.3 159 H Cl —OCH═CH₂ OH 7.6 681.3 683.3 160 OMe Br—O—CH(CH₃)₂ OH 7.5 771.2 773.2 161 H —SEt —OEt OH 7.0 709.3 162 H —SO₂Et—OEt OH 5.9 741.3 163 H

—OEt OH 7.1 735.2 164 H Cl

6.8 865.1 165 H Cl —SCH₂CH₂CH₃

9.5 816.3 166 H SEt —OEt

7.2 815.3

TABLE 2

(MH)⁺/ Cpd R³ R²² R²¹ R²⁰ R¹ t_(R) (MH + 2)⁺ 201 Et H F OEt OH 5.8 627.4202

H F OEt OH 5.6 645.4 203 —CH₂CH₂CH₃ H F OEt OH 6.3 641.4 204

H F OEt OH 6.5 695.4 205

H F OEt OH 5.2 669.4 206

H Cl OEt OH 6.5  661.4/ 663.4 207 —CH₂CH₂CH₃ H Cl OEt OH 6.9  657.4/659.4 208

H Cl OEt OH 6.9  711.4/ 713.4 209

H Cl OEt OH 6.1  685.4/ 687.4 210 Et H Me OEt OH 5.2 623.4 211

H Me OEt OH 4.9 641.4 212 —CH₂CH₂CH₃ H Me OEt OH 5.6 637.5 213

H Me OEt OH 5.8 691.4 214

H Me OEt OH 4.6 665.5 215

H Cl OEt OH 6.6  675.3/ 677.3 216

H Me OEt OH 5.0 665.4 217

H Me OEt

5.2 761.3

TABLE 3

Cpd R²² R²¹ R²⁰ R¹ t_(R) (MH)⁺ (MH + 2)⁺ 301 OMe Br OEt OH 7.6 759.2761.2 302 H Br OEt OH 8.1 729.2 731.2 303 H Cl OEt OH 7.9 685.3 687.3304 H F OEt OH 7.3 669.3 305 OMe Me OEt OH 6.4 695.4 306 H Me OEt OH 6.9665.3 307 H F OMe OH 7.2 655.3 308 OMe Me OMe OH 6.2 681.3 309 H Cl SEtOH 8.4 701.3 703.3 310 H CH₃ —OEt

8.5 804.4 311 OMe CH₃ —OEt

5.8 801.4

TABLE 4

Cpd R²² R²¹ R²³ t_(R) (MH)⁺ (MH + 2)⁺ 401 OMe Br H 6.4 729.2 731.2 402OMe Br Me 6.8 743.2 745.2 403 OMe Br Et 6.9 757.3 759.3 404 OMe Cl Me6.3 699.3 701.3 405 OMe Cl —CH₂CH═CH₂ 6.6 725.3 727.3 406 OMe CH₃ H 5.9665.3 667.3

TABLE 5

Cpd R²⁴ t_(R) (MH)⁺ 501 H 5.6 655.3 502 CH₃ 6.2 669.3

1. A compound of formula (I):

wherein R¹ is hydroxy; R² is a group of formula:

wherein R²⁰ is H, OH, halogen, or Y¹—R^(20a) wherein Y¹ is a bond, O, S,or NR^(20b) and wherein: R^(20a) is selected from the group consistingof: (C₁₋₈)alkyl, (C₁₋₆)alkyl-C≡N, (C₂₋₈)alkenyl, (C₂₋₈)alkynyl and(C₃₋₇)cycloalkyl, each of said alkyl, alkenyl, alkynyl and cycloalkylbeing optionally substituted with one, two or three substituents, eachindependently selected from: halogen, (C₁₋₆)alkyl optionally substitutedwith —O—(C₁₋₆)alkyl or —O—(C₃₋₆)cycloalkyl, (C₃₋₇)cycloalkyl,—O—(C₁₋₆)alkyl, Het, —O—(C₃₋₆)cycloalkyl, —NH₂, —NH(C₁₋₄)alkyl and—N((C₁₋₄)alkyl)₂; and R^(20b) is H, (C₁₋₆)alkyl or (C₃₋₆)cycloalkyl; andW is N; and the dotted line “a” is a double bond; or R²⁰ is oxo, and Wis NR²³ wherein R²³ is H, (C₁₋₆)alkyl, (C₂₋₆)alkenyl or (C₂₋₆)alkynyl;and the dotted line “a” is a single bond; R²¹ is halogen or Y²—R^(21a),wherein Y² is a bond, O, S, SO or SO₂, and R^(21a) is (C₁₋₆)alkyl,(C₂₋₆)alkenyl, (C₂₋₆)alkynyl, (C₃₋₇)cycloalkyl or(C₃₋₇)cycloalkyl-(C₁₋₆)alkyl-; R²² is H, —OH, —O—(C₁₋₄)alkyl, —NH₂,—NH(C₁₋₄)alkyl or —N((C₁₋₄)alkyl)₂; or R² is a group of formula:

wherein R²⁰, W and the dotted line “a” are as defined above; R²⁴ is H orR²¹ as defined above; and R²⁵ is H or (C₁₋₆)alkyl; X is O or NH; R³ is(C₁₋₁₀)alkyl, (C₃₋₇)cycloalkyl or (C₃₋₇)cycloalkyl-(C₁₋₄)alkyl-, a)wherein the cycloalkyl and cycloalkyl-alkyl- may be mono-, di- ortri-substituted with (C₁₋₃)alkyl; b) wherein the alkyl, cycloalkyl andcycloalkyl-alkyl- may be mono- or di-substituted with substituents eachindependently selected from hydroxy and O—(C₁₋₆)alkyl; c) wherein eachalkyl group may be mono-, di- or tri-substituted with halogen; and d)wherein in each cycloalkyl group being 5-, 6- or 7-membered, one or two—CH₂-groups not being directly linked to each other may be replaced by—O— such that the O-atom is linked to the group X via at least twoC-atoms; D is a 3 to 8 atom saturated or unsaturated alkylene chain; andthe dotted line “b” is a single bond or a double bond; wherein Het asused herein is defined as a 3- to 7-membered heterocycle having 1 to 4heteroatoms each independently selected from O, N and S, which may besaturated, unsaturated or aromatic, and which is optionally fused to atleast one other cycle to form a 4- to 14-membered heteropolycycle havingwherever possible 1 to 5 heteroatoms, each independently selected fromO, N and S, said heteropolycycle being saturated, unsaturated oraromatic; or a pharmaceutically acceptable salt or ester thereof.
 2. Thecompound according to claim 1 wherein R² is a group of formula:

wherein W, R²⁰, R²¹, R²² and the dotted line “a” are defined as inclaim
 1. 3. The compound according to claim 2 wherein R²⁰ is oxo; W isNR²³ wherein R²³ is Me, Et, —CH₂CH═CH₂ or H; and the dotted line “a” isa single bond.
 4. The compound according to claim 2 wherein W is N; thedotted line “a” is a double bond; and R²⁰ is H, OH, halogen, orY¹—R^(20a) wherein Y¹ is a bond, O, S, or NR^(20b); R^(20a) is selectedfrom the group consisting of: (C₁₋₈)alkyl, (C₂₋₈)alkenyl, (C₂₋₈)alkynyland (C₃₋₇)cycloalkyl, each of said alkyl, alkenyl, alkynyl andcycloalkyl being optionally substituted with one, two or threesubstituents, each independently selected from: halogen, (C₁₋₆)alkyl,(C₃₋₇)cycloalkyl, Het, —O—(C₁₋₆)alkyl, —O—(C₃₋₆)cycloalkyl, —NH₂,—NH(C₁₋₄)alkyl and —N((C₁₋₄)alkyl)₂; and R^(20b) is H, (C₁₋₆)alkyl or(C₃₋₆)cycloalkyl.
 5. The compound according to claim 2 wherein W is N;the dotted line “a” is a double bond; and R²⁰ is H, (C₁₋₆)alkyl, OH,—O—(C₁₋₆)alkyl, —S—(C₁₋₆)alkyl, —(CH₂)₀₋₄—CH═CH₂, —(CH₂)₀₋₄—C≡CH,—O—(CH₂)₀₋₄—CH═CH₂, —O—(CH₂)₀₋₄—C≡CH, —O—(CH₂)₁₋₄—OMe;—O—(CH₂)₁₋₄—N(Me)₂; —O—(CH₂)₁₋₄-Het; —S—(CH₂)₀₋₄—CH═CH₂,—S—(CH₂)₀₋₄—C≡CH, —S—(CH₂)₁₋₄—OMe; —S—(CH₂)₁₋₄—N(Me)₂, —S—(CH₂)₁₋₄-Het;(C₃₋₆)cycloalkyl, —O—(C₃₋₆)cycloalkyl, O—(C₁₋₆)alkyl-(C₃₋₆)cycloalkyl,—S—(C₃₋₆)cycloalkyl, or —S—(C₁₋₆)alkyl-(C₃₋₆)cycloalkyl; wherein Het is5- or 6-membered monocyclic heteroaryl containing from one to threeheteroatoms each independently selected from N, O and S; each of said(C₁₋₆)alkyl, —O—(C₁₋₆)alkyl, —S—(C₁₋₆)alkyl, —(CH₂)₀₋₄—CH═CH₂,—(CH₂)₀₋₄—C≡CH, —O—(CH₂)₀₋₄—CH═CH₂, —O—(CH₂)₀₋₄—C≡CH,—S—(CH₂)₀₋₄—CH═CH₂, —S—(CH₂)₀₋₄—C≡CH, (C₃₋₆)cycloalkyl,—O—(C₃₋₆)cycloalkyl, and —S—(C₃₋₆)cycloalkyl being optionallysubstituted with one, two or three substituents, each independentlyselected from (C₁₋₄)alkyl, —O—(C₁₋₄)alkyl, and halo; or R²⁰ isNR^(20a)R^(20b) wherein R^(20a) is (C₁₋₄)alkyl, and R^(20b) is H,(C₁₋₄)alkyl or (C₃₋₅)cycloalkyl.
 6. The compound according to claim 5wherein R²⁰ is H, methyl, ethyl, propyl, 1-methylethyl, butyl,2-methylpropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,—CH═CH₂, —C≡CH, O-methyl, O-ethyl, O-propyl, O—CH(CH₃)₂, 0-cyclopropyl,O-cyclobutyl, O-cyclopentyl, O-cyclohexyl, O—CH₂CH₂CF₃, —O—CH═CH₂,—O—CH₂—CH═CH₂, O—C≡CH, —O—CH₂—C≡CH, —O—CH₂—C≡CCH₃, —O—CH₂—CH₂—OMe,—O—CH₂—CH₂—N(Me)₂,S-methyl, S-ethyl, S-propyl, S—CH(CH₃)₂,S-cyclopropyl, S-cyclobutyl, S-cyclopentyl, S-cyclohexyl, —S—CH═CH₂,—S—CH₂—CH═CH₂, S—C≡CH, —S—CH₂—C≡CH, —S—CH₂—CH₂—OMe, —S—CH₂—CH₂—N(Me)₂,


7. The compound according to claim 2 wherein R²¹ is selected from:fluorine, chlorine, bromine, —CH₃, —CH₂CH₃, —CH₂CH₂CH₃, —OCH₃, —OCH₂CH₃,—OCH₂CH₂CH₃, —SCH₃, —SCH₂CH₃, —SCH₂CH₂CH₃, —(SO)CH₃, —(SO)CH₂CH₃,—(SO)CH₂CH₂CH₃, —(SO₂)CH₃, —(SO₂)CH₂CH₃, —(SO₂)CH₂CH₂CH₃,

and —C≡CH.
 8. The compound according to claim 2 wherein R²² is selectedfrom: H, —OH, —OCH₃, —OCH₂CH₃, —OCH₂CH₂CH₃, —NHCH₃, —N(CH₃)₂,—N(CH₃)CH₂CH₃ and —N(CH₃)CH₂CH₂CH₃.
 9. The compound according to claim 8wherein R²² is H or —OCH₃.
 10. The compound according to claim 1 whereinR² is a group of formula:

wherein W, R²⁰, R²⁴, R²⁵ and the dotted line “a” are defined as inclaim
 1. 11. The compound according to claim 10 wherein W is N; thedotted line “a” is a double bond; and R²⁰ is H, OH, halogen, orY¹—R^(20a) wherein Y¹ is a bond, O, S, or NR^(20b); R^(20a) is selectedfrom the group consisting of: (C₁₋₈)alkyl, (C₂₋₈)alkenyl, (C₂₋₈)alkynyland (C₃₋₇)cycloalkyl, each of said alkyl, alkenyl, alkynyl andcycloalkyl being optionally substituted with one, two or threesubstituents, each independently selected from: halogen, (C₁₋₆)alkyl,(C₃₋₇)cycloalkyl, Het, —O—(C₁₋₆)alkyl, —O—(C₃₋₆)cycloalkyl, —NH₂,—NH(C₁₋₄)alkyl and —N((C₁₋₄)alkyl)₂; and R^(20b) is H, (C₁₋₆)alkyl or(C₃₋₆)cycloalkyl.
 12. The compound according to claim 11 wherein R²⁰ isY¹—R^(20a), wherein Y¹ is 0 and R^(20a) is (C₁₋₈)alkyl.
 13. The compoundaccording to claim 10 wherein R²⁴ is H or (C₁₋₆)alkyl.
 14. The compoundaccording to claim 10 wherein R²⁵ is H.
 15. The compound according toclaim 1 wherein X is O.
 16. The compound according to claim 1 wherein R³is selected from (C₂₋₈)alkyl, (C₃₋₇)cycloalkyl and(C₃₋₇)cycloalkyl-(C₁₋₃)alkyl-, a) wherein said cycloalkyl andcycloalkyl-alkyl- may be mono-, di- or tri-substituted with (C₁₋₃)alkyl;and b) wherein said alkyl, cycloalkyl and cycloalkyl-alkyl- may be mono-or di-substituted with substituents each independently selected fromhydroxy and O—(C₁₋₄)alkyl; and c) wherein each of said alkyl groups maybe mono-, di- or tri-substituted with fluorine or mono-substituted withchlorine or bromine; and d) wherein in each of said cycloalkyl groupsbeing 5-, 6- or 7-membered, one or two —CH₂-groups not being directlylinked to each other may be replaced by —O— such that the O-atom islinked to the group X via at least two C-atoms.
 17. The compoundaccording to claim 16 wherein R³ is selected from cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, and a group selected from:


18. The compound according to claim 1 wherein linker D is a 5 carbonatom chain.
 19. The compound according to claim 1 wherein the dottedline “b” is a single bond or a double bond in the Z (cis) configuration.20. The compound according to claim 1 of formula (Ia):

wherein R¹ is hydroxy; R²⁰ is H, OH, halogen, or Y¹—R^(20a) wherein Y¹is a bond, O, S, or NR^(20b) wherein: R^(20a) is selected from the groupconsisting of: (C₁₋₈)alkyl, (C₁₋₆)alkyl-C≡N, (C₂₋₈)alkenyl,(C₂₋₈)alkynyl, all of said alkyl, alkenyl and alkynyl being optionallymono- or di-substituted with: halogen, (C₁₋₆)alkyl, —O—(C₁₋₆)alkyl,(C₁₋₄)alkyl-O—(C₁₋₆)alkyl, —O—(C₃₋₆)cycloalkyl,(C₁₋₄)alkyl-O—(C₃₋₆)cycloalkyl, amino, (C₁₋₄)alkylamino, ordi((C₁₋₄)alkyl)amino; and R^(20b) is H, (C₁₋₆)alkyl or (C₃₋₆)cycloalkyl;and W is N; and the dotted line “a” is a double bond; or R²⁰ is oxo, andW is NR²³ wherein R²³ is H, (C₁₋₆)alkyl, (C₂₋₆)alkenyl, (C₂₋₆)alkynyl;and the dotted line “a” is a single bond; R²¹ is halogen, (C₁₋₆)alkyl,(C₂₋₆)alkenyl, (C₂₋₆)alkynyl, —O—(C₁₋₆)alkyl, —O—(C₂₋₆)alkenyl,—O—(C₂₋₆)alkynyl, —S—(C₁₋₆)alkyl, —S—(C₂₋₆)alkenyl, and—S—(C₂₋₆)alkynyl, wherein the sulfur is in any oxidized state; R²² is H,—OH, —O—(C₁₋₄)alkyl, —NH₂, —NH(C₁₋₄)alkyl or —N((C₁₋₄)alkyl)₂; X is O orNH; R³ is (C₁₋₁₀)alkyl, (C₃₋₇)cycloalkyl, (C₃₋₇)cycloalkyl-(C₁₋₄)alkyl-,a) wherein the cycloalkyl and cycloalkyl-alkyl- may be mono-, di- ortri-substituted with (C₁₋₃)alkyl; b) wherein the alkyl, cycloalkyl andcycloalkyl-alkyl- may be mono- or di-substituted with substituentsselected from hydroxy and O—(C₁₋₆)alkyl; c) wherein all the alkyl groupsmay be mono-, di- or tri-substituted with halogen; and d) wherein in thecycloalkyl groups, being 5-, 6- or 7-membered, one or two —CH₂-groupsnot being directly linked to each other may be replaced by —O—; D is a 3to 8 atom saturated or unsaturated alkylene chain; and the dotted line“b” is a single bond or a double bond; or a pharmaceutically acceptablesalt or ester thereof.
 21. The compound according to claim 1 wherein R¹is hydroxy; R² is a group of formula

wherein R²⁰ is oxo, W is NR²³ wherein R²³ is Me, Et, —CH₂CH═CH₂ or H andthe dotted line “a” is a single bond; or W is N, the dotted line “a” isa double bond; and R²⁰ is H, OH, halogen, or Y¹—R^(20a) wherein Y¹ is abond, O, S, or NR^(20b); R^(20a) is selected from the group consistingof: (C₁₋₈)alkyl, (C₂₋₈)alkenyl, (C₂₋₈)alkynyl and (C₃₋₇)cycloalkyl, eachof said alkyl, alkenyl, alkynyl and cycloalkyl being optionallysubstituted with one, two or three substituents, each independentlyselected from: halogen, (C₁₋₆)alkyl, (C₃₋₇)cycloalkyl, Het,—O—(C₁₋₆)alkyl, —O—(C₃₋₆)cycloalkyl, —NH₂, —NH(C₁₋₄)alkyl and—N((C₁₋₄)alkyl)₂; wherein Het is defined as in claim 1; and R^(20b) isH, (C₁₋₆)alkyl or (C₃₋₆)cycloalkyl; and R²¹ is selected from: fluorine,chlorine, bromine, —CH₃, —CH₂CH₃, —CH₂CH₂CH₃, —OCH₃, —OCH₂CH₃,—OCH₂CH₂CH₃, —SCH₃, —SCH₂CH₃, —SCH₂CH₂CH₃, —(SO)CH₃, —(SO)CH₂CH₃,—(SO)CH₂CH₂CH₃, —(SO₂)CH₃, —(SO₂)CH₂CH₃, —(SO₂)CH₂CH₂CH₃,

and —C≡CH; and R²² is selected from: H, —OH, —OCH₃, —OCH₂CH₃,—OCH₂CH₂CH₃, —NHCH₃, —N(CH₃)₂, —N(CH₃)CH₂CH₃ and —N(CH₃)CH₂CH₂CH₃; or R²is a group of formula

wherein W is N; the dotted line “a” is a double bond; and R²⁰ is H, OH,halogen, or Y¹—R^(20a) wherein Y¹ is a bond, O, S, or NR^(20b); R^(20a)is selected from the group consisting of: (C₁₋₈)alkyl, (C₂₋₈)alkenyl,(C₂₋₈)alkynyl and (C₃₋₇)cycloalkyl, each of said alkyl, alkenyl, alkynyland cycloalkyl being optionally substituted with one, two or threesubstituents, each independently selected from: halogen, (C₁₋₆)alkyl,(C₃₋₇)cycloalkyl, Het, —O—(C₁₋₆)alkyl, —O—(C₃₋₆)cycloalkyl, —NH₂,—NH(C₁₋₄)alkyl and —N((C₁₋₄)alkyl)₂; wherein Het is defined as in claim1; and R^(20b) is H, (C₁₋₆)alkyl or (C₃₋₆)cycloalkyl; and R²⁴ is H or(C₁₋₆)alkyl; and R²⁵ is H; and X is O or NH; and R³ is selected from(C₂₋₈)alkyl, (C₃₋₇)cycloalkyl and (C₃₋₇)cycloalkyl-(C₁₋₃)alkyl-, a)wherein said cycloalkyl and cycloalkyl-alkyl- may be mono-, di- ortri-substituted with (C₁₋₃)alkyl; and b) wherein said alkyl, cycloalkyland cycloalkyl-alkyl- may be mono- or di-substituted with substituentseach independently selected from hydroxy and O—(C₁₋₄)alkyl; and c)wherein each of said alkyl groups may be mono-, di- or tri-substitutedwith fluorine or mono-substituted with chlorine or bromine; and d)wherein in each of said cycloalkyl groups being 5-, 6- or 7-membered,one or two —CH₂-groups not being directly linked to each other may bereplaced by —O— such that the O-atom is linked to the group X via atleast two C-atoms; and linker D is a 3 to 8 atom saturated orunsaturated alkylene chain; and the dotted line “b” is a single bond ora double bond.
 22. The compound according to claim 1 wherein R¹ ishydroxy; R² is a group of formula

wherein R²⁰ is oxo, W is NR²³ wherein R²³ is Me, Et, —CH₂CH═CH₂ or H andthe dotted line “a” is a single bond; or W is N, the dotted line “a” isa double bond, and R²⁰ is H, (C₁₋₆)alkyl, OH, —O—(C₁₋₆)alkyl,—S—(C₁₋₆)alkyl, —(CH₂)₀₋₄—CH═CH₂, —(CH₂)₀₋₄—C≡CH, —O—(CH₂)₀₋₄—CH═CH₂,—O—(CH₂)₀₋₄—C≡CH, —O—(CH₂)₁₋₄—OMe; —O—(CH₂)₁₋₄—N(Me)₂; —O—(CH₂)₁₋₄-Het;—S—(CH₂)₀₋₄—CH═CH₂, —S—(CH₂)₀₋₄—C≡CH, —S—(CH₂)₁₋₄—OMe;—S—(CH₂)₁₋₄—N(Me)₂, —S—(CH₂)₁₋₄-Het; (C₃₋₆)cycloalkyl,—O—(C₃₋₆)cycloalkyl, O—(C₁₋₆)alkyl-(C₃₋₆)cycloalkyl,—S—(C₃₋₆)cycloalkyl, or —S—(C₁₋₆)alkyl-(C₃₋₆)cycloalkyl; wherein Het is5- or 6-membered monocyclic heteroaryl containing from one to threeheteroatoms each independently selected from N, O and S; each of said(C₁₋₆)alkyl, —O—(C₁₋₆)alkyl, —S—(C₁₋₆)alkyl, —(CH₂)₀₋₄—CH═CH₂,—(CH₂)₀₋₄—C≡CH, —O—(CH₂)₀₋₄—CH═CH₂, —O—(CH₂)₀₋₄—C≡CH,—S—(CH₂)₀₋₄—CH═CH₂, —S—(CH₂)₀₋₄—C≡CH, (C₃₋₆)cycloalkyl,—O—(C₃₋₆)cycloalkyl, and —S—(C₃₋₆)cycloalkyl being optionallysubstituted with one, two or three substituents, each independentlyselected from (C₁₋₄)alkyl, —O—(C₁₋₄)alkyl, and halo; or R²⁰ isNR^(20a)R^(20b) wherein R^(20a) is (C₁₋₄)alkyl, and R²⁰b is H,(C₁₋₄)alkyl or (C₃₋₅)cycloalkyl; and R²¹ is selected from: fluorine,chlorine, bromine, —CH₃, —CH₂CH₃, —CH₂CH₂CH₃, —OCH₃, —OCH₂CH₃,—OCH₂CH₂CH₃, —SCH₃, —SCH₂CH₃, —SCH₂CH₂CH₃, —(SO)CH₃, —(SO)CH₂CH₃,—(SO)CH₂CH₂CH₃, —(SO₂)CH₃, —(SO₂)CH₂CH₃, —(SO₂)CH₂CH₂CH₃,

 and —C≡CH; and R²² is selected from H, —OCH₃ and —N(CH₃)₂; or R² is agroup of formula

wherein W is N; the dotted line “a” is a double bond; R²⁰ is Y¹—R^(20a),wherein Y¹ is O and R^(20a) is (C₁₋₈)alkyl; R²⁴ is H or (C₁₋₆)alkyl; andR²⁵ is H; and X is O or NH; and R³ is selected from cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, and a group selected from:

and linker D is a 3 to 8 atom saturated or unsaturated alkylene chain;and the dotted line “b” is a single bond or a double bond.
 23. Thecompound according to claim 1 wherein R¹ is hydroxy; R² is a group offormula

wherein R²⁰ is oxo, W is NR²³ wherein R²³ is Me, Et, —CH₂CH═CH₂ or H andthe dotted line “a” is a single bond; or W is N, the dotted line “a” isa double bond, and R²⁰ is H, methyl, ethyl, 1-methylethyl, —C≡CH,O-methyl, O-ethyl, O— propyl, O—CH(CH₃)₂, O-cyclopentyl, O—CH₂CH₂CF₃,—O—CH═CH₂, —O—CH₂—CH═CH₂, —O—CH₂—C≡CH, —O—CH₂—C≡CCH₃, —O—CH₂CH₂OMe,—O—CH₉CH₉N(Me)₂, S-methyl, S-ethyl, S-propyl, S—CH(CH₃)₂,

and R²¹ is selected from fluorine, chlorine, bromine, —CH₃, —OCH₃,—SCH₃, —SCH₂CH₃, (SO)CH₃, (SO₂)CH₃, —(SO₂)CH₂CH₃,

 and —C≡CH; and R²² is H or —OCH₃; or R² is a group of formula

wherein W is N; the dotted line “a” is a double bond; R²⁰ is —O—CH₂CH₃;R²⁴ is H or CH₃; and R²⁵ is H; and X is O; and R³ is selected fromcyclopentyl,

and linker D is a 5 carbon atom chain; and the dotted line “b” is asingle bond or a double bond in the Z (cis) configuration.
 24. Thecompound according to claim 1 of the formula:

wherein R¹, R²⁰, R²¹ and R²² are defined as in the table below: Cpd R²²R²¹ R²⁰ R¹ 101 OMe Br OEt OH 102 OMe Me OEt OH 103 H Br OEt OH 104 H ClOEt OH 105 H Br OMe OH 106 H Cl OMe OH 107 H F OMe OH 108 OMe Me OMe OH109 OMe Me H OH 110 H Cl H OH 111 H Br H OH 112 H Cl Me OH 113 H F OEtOH 114 H Me OEt OH 115 H Br Me OH 116 H Cl Et OH 117 H Cl —CH(CH₃)₂ OH118 H SMe OEt OH 119 H Me O—CH(CH₃)₂ OH 120 OMe Me O—CH(CH₃)₂ OH 121 HCl O—CH(CH₃)₂ OH 122 H Br O—CH(CH₃)₂ OH 123 H F O—CH(CH₃)₂ OH 124 HSO₂ME OEt OH 125 H Cl SMe OH 126 OMe Br OMe OH 127 H SMe H OH 128 H SOMeH OH 129 H SO₂Me H OH 130 H Cl SEt OH 131 H Cl OCH₂CH(CH₃)₂ OH 132 H ClOCH₂CH₂CH₃ OH 133 H OMe OEt OH 134 H —C≡CH OEt OH 137 H Cl SCH₂CH₂CH₃ OH138 H Cl SCH(CH₃)₂ OH 139 H Cl OCH₂C(CH₃)₃ OH 140 H Cl OCH₂CH₂CF₃ OH 141H Cl

OH 142 H Cl

OH 143 H Cl

OH 144 H Cl

OH 145 H Cl —C≡CH OH 146 OMe Br —OCH₂C≡CH OH 147 OMe Br —OCH₂CH═CH₂ OH148 H Cl

OH 149 OMe CH₃ —OCH₂CH═CH₂ OH 150 OMe CH₃ —OCH₂C≡CH OH 151 OMe Cl —OEtOH 152 OMe Cl —O—CH(CH₃)₂ OH 153 OMe Cl —OMe OH 154 OMe Cl —OCH₂CH≡CH₂OH 155 OMe Cl —OCH₂C≡CH OH 156 OMe Br —OCH₂C≡CCH₃ OH 157 H Cl

OH 158 H Cl

OH 159 H Cl —OCH═CH₂ OH 160 OMe Br —O—CH(CH₃)₂ OH 161 H —SEt —OEt OH 162H —SO₂Et —OEt OH 163 H

—OEt OH


25. The compound according to claim 1 of the formula:

wherein R¹, R²⁰, R²¹, R²² and R³ are defined as in the table below: CpdR³ R²² R²¹ R²⁰ R¹ 201 Et H F OEt OH 202

H F OEt OH 203 —CH₂CH₂CH₃ H F OEt OH 204

H F OEt OH 205

H F OEt OH 206

H Cl OEt OH 207 —CH₂CH₂CH₃ H Cl OEt OH 208

H Cl OEt OH 209

H Cl OEt OH 210 Et H Me OEt OH 211

H Me OEt OH 212 —CH₂CH₂CH₃ H Me OEt OH 213

H Me OEt OH 214

H Me OEt OH 215

H Cl OEt OH 216

H Me OEt OH


26. The compound according to claim 1 of the formula:

wherein R¹, R²⁰, R²¹ and R²² are defined as in the table below: Cpd R²²R²¹ R²⁰ R¹ 301 OMe Br OEt OH 302 H Br OEt OH 303 H Cl OEt OH 304 H F OEtOH 305 OMe Me OEt OH 306 H Me OEt OH 307 H F OMe OH 308 OMe Me OMe OH309 H Cl SEt OH


27. The compound according to claim 1 of the formula:

wherein R²¹, R²² and R²³ are defined as in the table below: Cpd R²² R²¹R²³ 401 OMe Br H 402 OMe Br Me 403 OMe Br Et 404 OMe Cl Me 405 OMe Cl—CH₂CH═CH₂ 406 OMe CH₃ H


28. The compound according to claim 1 of the formula:

wherein R²⁴ is defined as in the table below: Cpd R²⁴ 501 H 502 CH₃


29. A pharmaceutical composition comprising an anti-hepatitis C virallyeffective amount of a compound according to claim 1 or apharmaceutically acceptable salt or ester thereof, and apharmaceutically acceptable carrier medium or auxiliary agent.
 30. Thepharmaceutical composition according to claim 29 further comprising atherapeutically effective amount of at least one other antiviral agent.31. The pharmaceutical composition according to claim 30, wherein saidantiviral agent is ribavirin.
 32. The pharmaceutical compositionaccording to claim 30, wherein said antiviral agent is selected from another anti-HCV agent, HIV inhibitor, HAV inhibitor and HBV inhibitor.33. The pharmaceutical composition according to claim 32, wherein saidother anti-HCV agent is selected from the group consisting ofimmunomodulatory agents, other inhibitors of HCV NS3 protease,inhibitors of HCV polymerase and inhibitors of another target in the HCVlife cycle.
 34. The pharmaceutical composition according to claim 33,wherein said immunomodulatory agent is selected from α-interferon,γ-interferon and pegylated α-interferon.
 35. The pharmaceuticalcomposition according to claim 33, wherein said inhibitor of anothertarget in the HCV life cycle is selected from inhibitors of: helicase,NS2/3 protease and internal ribosome entry site (IRES).
 36. A method forthe treatment or prevention of a hepatitis C viral infection in a mammalcomprising administering to the mammal an anti-hepatitis C virallyeffective amount of a compound according to claim 1, or apharmaceutically acceptable salt or ester thereof.
 37. A method for thetreatment or prevention of a hepatitis C viral infection in a mammalcomprising administering to the mammal an anti-hepatitis C virallyeffective amount of a combination of a compound according to claim 1, ora pharmaceutically acceptable salt or ester thereof, and at least oneother antiviral agent.
 38. The method according to claim 37, whereinsaid antiviral agent is ribavirin.
 39. The method according to claim 37wherein said other antiviral agent is selected from another anti-HCVagent, HIV inhibitor, HAV inhibitor and HBV inhibitor.
 40. The methodaccording to claim 39, wherein said other anti-HCV agent is selectedfrom immunomodulatory agents, other inhibitors of HCV NS3 protease,inhibitors of HCV polymerase and inhibitors of another target in the HCVlife cycle.
 41. The method according to claim 40, wherein saidimmunomodulatory agent is selected from α-interferon, γ-interferon andpegylated α-interferon.
 42. The method according to claim 40, whereinsaid inhibitor of another target in the HCV life cycle is selected frominhibitors of: helicase, NS2/3 protease and internal ribosome entry site(IRES).
 43. A method of inhibiting the replication of hepatitis C virusby exposing the virus to a hepatitis C viral NS3 protease inhibitingamount of a compound according to claim 1, or a pharmaceuticallyacceptable salt or ester thereof.
 44. An article of manufacturecomprising a composition effective to treat an HCV infection or toinhibit the NS3 protease of HCV and packaging material comprising alabel which indicates that the composition can be used to treatinfection by the hepatitis C virus, wherein said composition comprises acompound according to claim 1 or a pharmaceutically acceptable salt orester thereof.